U.S. patent application number 15/894652 was filed with the patent office on 2019-01-24 for pharmaceutical formulations containing microparticles or nanoparticles of a delivery agent.
The applicant listed for this patent is Emisphere Technologies, Inc.. Invention is credited to Ehud Arbit, Nikhil Dhoot, Steven Dinh, Jamila Harris, George Klein, Jongbin Lee, Halina Levchik, Jun Liao, Puchun Liu, Shingai Majuru, Nai Fang Wang.
Application Number | 20190022228 15/894652 |
Document ID | / |
Family ID | 37431701 |
Filed Date | 2019-01-24 |
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United States Patent
Application |
20190022228 |
Kind Code |
A1 |
Klein; George ; et
al. |
January 24, 2019 |
PHARMACEUTICAL FORMULATIONS CONTAINING MICROPARTICLES OR
NANOPARTICLES OF A DELIVERY AGENT
Abstract
This invention relates to microparticles and/or nanoparticles
containing a delivery agent and/or an active agent. This invention
also relates to pharmaceutical formulations and solid dosage forms,
including controlled release solid dosage forms of active agent and
a delivery agent.
Inventors: |
Klein; George; (Tarrytown,
NY) ; Majuru; Shingai; (Greensboro, NC) ; Liu;
Puchun; (Chappaqua, NY) ; Dinh; Steven; (Coral
Gables, FL) ; Liao; Jun; (Roseland, NJ) ; Lee;
Jongbin; (New City, NY) ; Levchik; Halina;
(Croton On Hudson, NY) ; Arbit; Ehud; (Englewood,
NJ) ; Dhoot; Nikhil; (Dombivli (east), IN) ;
Harris; Jamila; (Flushing, NY) ; Wang; Nai Fang;
(Long Island City, NY) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Emisphere Technologies, Inc. |
Roseland |
NJ |
US |
|
|
Family ID: |
37431701 |
Appl. No.: |
15/894652 |
Filed: |
February 12, 2018 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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12550281 |
Aug 28, 2009 |
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15894652 |
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11204756 |
Aug 15, 2005 |
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12550281 |
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60612810 |
Sep 23, 2004 |
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60601258 |
Aug 13, 2004 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 9/2013 20130101;
A61K 9/2846 20130101; A61K 9/145 20130101; A61K 9/4858 20130101;
A61K 47/12 20130101; A61K 9/2027 20130101; A61K 38/28 20130101;
A61K 9/2081 20130101; A61P 3/10 20180101; A61P 43/00 20180101; A61K
9/1617 20130101; A61P 19/08 20180101; A61K 9/2077 20130101; A61P
5/50 20180101 |
International
Class: |
A61K 47/12 20060101
A61K047/12; A61K 38/28 20060101 A61K038/28; A61K 9/48 20060101
A61K009/48; A61K 9/28 20060101 A61K009/28; A61K 9/14 20060101
A61K009/14; A61K 9/20 20060101 A61K009/20; A61K 9/16 20060101
A61K009/16 |
Claims
1-11. (canceled)
12. A pharmaceutical formulation comprising particles having a
median particle size of less than about 999 micrometers, the
particles comprising a delivery agent and an active agent.
13-16. (canceled)
17. The pharmaceutical formulation of claim 12, wherein the
particles have a median particle size of about 45 to about 150
micrometers.
18. The pharmaceutical formulation of claim 12, wherein the
particles have a median particle size of about 150 to about 250
micrometers.
19. The pharmaceutical formulation of claim 12, wherein the
particles have a median particle size of about 250 to about 425
micrometers.
20. The pharmaceutical formulation of claim 12, wherein the
particles have a median particle size of about 425 to about 850
micrometers.
21. The pharmaceutical formulation of claim 12, wherein the
particles have a median particle size of about 100 to about 1000
nanometers.
22. The pharmaceutical formulation of claim 21, wherein the
particles have a median particle size of about 500 to about 1000
nanometers.
23-38. (canceled)
39. The pharmaceutical formulation of claim 12, wherein the
delivery agent compound is selected from
N-(8-[2-hydroxybenzoyl]amino)caprylic acid,
N-(10-[2-hydroxybenzoyl]-amino)decanoic acid,
8-(2-hydroxy-4-methoxybenzoylamino)octanoic acid,
8-(2-hydroxy-5-chlorobenzoylamino)-octanoic acid,
4-[(2-hydroxy-4-chlorobenzoyl)-amino]butanoic acid, and
pharmaceutically acceptable salts thereof.
40. The pharmaceutical formulation of claim 12, wherein the
delivery agent compound is N-(8-[2-hydroxybenzoyl]-amino)caprylic
acid or a pharmaceutically acceptable salt thereof.
41. The pharmaceutical formulation of claim 12, wherein the
delivery agent compound is N-(10-[2-hydroxybenzoyl]-amino)decanoic
acid or a pharmaceutically acceptable salt thereof.
42. The pharmaceutical formulation of claim 12, wherein the
delivery agent compound is
4-[(2-hydroxy-4-chloro-benzoyl)-amino]butanoic acid or a
pharmaceutically acceptable salt thereof.
43. The pharmaceutical formulation of claim 12, wherein the active
agent is selected from proteins, polypeptides, peptides, hormones,
and polysaccharides.
44. The pharmaceutical formulation of claim 12, wherein the active
agent is selected from the following, including synthetic, natural
or recombinant sources thereof: growth hormones; growth hormone
releasing hormones; growth hormone releasing factor, interferons;
interleukin-1; interleukin-2; insulin, optionally having counter
ions including zinc, sodium, calcium and ammonium; insulin-like
growth factor; heparin; calcitonin; erythropoietin; atrial
naturetic factor; antigens; monoclonal antibodies; somatostatin;
protease inhibitors; adrenocorticotropin, gonadotropin releasing
hormone; oxytocin; leutinizing-hormone-releasing-hormone; follicle
stimulating hormone; glucocerebrosidase; thrombopoietin;
filgrastim; prostaglandins; cyclosporin; vasopressin; cromolyn
sodium; vancomycin; desferrioxamine; bisphosphonates; parathyroid
hormone; anti-migraine agents; glucagon-like peptide 1 (GLP-1);
antimicrobials; vitamins; analogs, fragments, mimetics or
polyethylene glycol (PEG)-modified derivatives of these compounds;
or any combination thereof.
45. The pharmaceutical formulation of claim 12, wherein the active
agent is insulin.
46-49. (canceled)
50. A solid dosage unit form comprising the pharmaceutical
formulation of claim 12.
51-56. (canceled)
57. A method of treating diabetes in a mammal in need thereof,
comprising administering to the animal a therapeutic effective
amount of a pharmaceutical formulation of claim 12.
58. The method of claim 57, wherein the delivery agent compound is
4-[(2-hydroxy-4-chloro-benzoyl)-amino]butanoic acid or a
pharmaceutically acceptable salt thereof.
59. A method of treating impaired glucose tolerance, early stage
diabetes, or late stage diabetes or achieving glucose homeostasis
in humans, comprising administering a therapeutic effective amount
of a pharmaceutical formulation of claim 12.
60. (canceled)
61. A method of treating a human diabetic patient comprising orally
administering to the human diabetic patient on a chronic basis a
therapeutic effective amount of a pharmaceutical formulation of
claim 12.
62-94. (canceled)
Description
[0001] This application is a continuation of U.S. application Ser.
No. 11/204,756, filed Aug. 15, 2005, and claims the benefit of U.S.
Provisional Application No. 60/612,810, filed Sep. 23, 2004, and
U.S. Provisional Application No. 60/601,258, filed Aug. 13, 2004,
each of which are hereby incorporated by reference.
FIELD OF THE INVENTION
[0002] This invention relates to pharmaceutical formulations and
methods for preparing the same.
BACKGROUND OF THE INVENTION
[0003] There is a continuing need for improved oral delivery
systems for drugs, such as insulin.
SUMMARY OF THE INVENTION
[0004] The present invention relates to microparticles and/or
nanoparticles for oral administration containing a delivery agent
compound alone or a combination of a delivery agent compound and an
active agent. Formulations containing these particles (and, for
particles containing only a delivery agent compound, and an active
agent) provide significantly greater bioavailability of the active
agent with less variability than oral administration of a simple
mixture of the delivery agent compound and active agent as a
powder, tablet, or capsule. Without being bound by any particular
theory, it is believed that in at least some embodiments, this
improvement may be due to (1) the small size of the micro- or
nano-particles which permits them to pass from the stomach, through
the pylorus (which typically has a diameter of 1000-2000 .mu.m), to
the small intestine, where particle dissolution and delivery
agent-mediated drug absorption is believed to best occur, and (2)
the intimate contact between the delivery agent compound and active
agent in the particles which ensures that the delivery agent
compound is present with the active agent at the site of
absorption. Because the micro- and nano-particles freely pass
through the pylorus into the small intestine, unlike a conventional
tablet or capsule which must first become dissolved into particles
sufficiently small to do so, variations caused by tablet
disintegration and gastric transit modulated by gastric motility
are minimized.
[0005] According to one embodiment, the particles comprising a
delivery agent compound and an active agent have a median particle
size less than about 900 or 1000 .mu.m. For example, the median
particle size can range from about 45 to about 850 .mu.m, from
about 45 to about 150 .mu.m, from about 150 to about 250 .mu.m,
from about 250 to about 425 .mu.m, from about 425 to about 850
.mu.m, from about 100 to about 1000 nm, or from about 500 to about
1000 nm. According to another embodiment, the particles have a
median particle size less than about 1 .mu.m. In some embodiments,
particles may be as small as about 1 nanometer and as large as
about 999 micrometers. For example, the particles may have a median
particle size of less than about 999 micrometers, from about 1
nanometer to about 999 micrometers, about 1 to about 999
micrometers, about 1 to about 999 nanometers, about 45 to about 850
micrometers, about 45 to about 150 micrometers, about 150 to about
250 micrometers, about 250 to about 425 micrometers, about 425 to
about 850 micrometers, about 100 to about 1000 nanometers, or about
500 to about 1000 nanometers.
[0006] Another embodiment is a pharmaceutical formulation
comprising a delivery agent compound and an active agent in which
the delivery agent compound is in the form of particles. The
particles can have a median particle size of less than about 999
micrometers, about 1 nanometer to about 999 micrometers, about 1 to
about 999 nanometers, or about 7 to about 16 micrometers.
Optionally, the active agent may also be in the form of particles.
For example, the median particle size of the active agent particles
may be less than about 999 micrometers, about 1 nanometer to about
999 micrometers, about 1 to about 999 micrometers, or about 1 to
about 999 nanometers. According to one embodiment, the delivery
agent particles and the active agent particles both have a median
particle size of about 1 to about 999 micrometers. According to
another embodiment, the delivery agent particles and the active
agent particles both have a median particle size of about 1 to
about 999 nanometers.
[0007] Yet another embodiment is a pharmaceutical formulation
comprising a delivery agent and an active agent in which the active
agent is in the form of particles having a median particle size of
less than about 999 micrometers. According to one embodiment, the
median particle size of the active agent particles is about 1
nanometer to about 999 micrometers, about 1 to about 999
micrometers, or about 1 to about 999 nanometers.
[0008] The particles can be in the form of fine granules or
micro-beads (e.g., beads having a round/ball shape and a diameter
of about 0.2 mm to about 2.0 mm). The micro-beads may be formed by
compression. In one embodiment, the pharmaceutical formulation
includes micro-beads containing a delivery agent compound, which
are coated with an active agent, such as insulin or heparin. The
micro-beads may have a diameter ranging from about 0.2 mm to 2.0
mm.
[0009] The particles may also include a mucoadhesive, such as a
cellulose derivative (e.g., CMC sodium (available from Aqualon of
Wilmington, Del.)) or a polyacrylic acid (e.g., Carbopol.TM.
available from B.F. Goodrich of Cleveland, Ohio). The mucoadhesive
can (1) facilitate adhesion to mucosa (including in the
gastrointestinal tract) thereby prolonging delivery agent-active
agent contact with the mucosa, (2) stabilize and protect the active
agent (e.g., in the case of insulin), and (3) increase the
permeability of biomembranes (including mucosa) thereby improving
delivery and increasing bioavailability of the active agent.
[0010] It has also been discovered that oral administration of
insulin in conjunction with a delivery agent compound by solid oral
dosage forms that do not degrade in the stomach, but do degrade in
the intestine, provides significantly greater bioavailability of
the insulin. Such solid oral dosage forms containing insulin or a
different active agent provide greater bioavailability than forms
that degrade in the stomach and forms that do not contain the
delivery agent compound. Without being bound by any particular
theory, it is believed that this improvement is due to the
sensitivity of insulin and other active agents to degradation by
enzymes or acid found in gastric fluid. Because the solid oral
dosage forms do not degrade in the stomach, the insulin and other
active agents are protected from degradation until they reach the
intestine.
[0011] Another embodiment of the invention is a pharmaceutical
formulation (such as a solid oral dosage form) comprising a
therapeutically effective amount of an active agent and a delivery
agent, where the pharmaceutical formulation has a disintegration
time of about 250 seconds to about 650 seconds when orally
administered. In another embodiment, the disintegration time is
about 350 to about 550 seconds when orally administered. In yet
another embodiment, the disintegration time is greater than 60
seconds when orally administered. In yet another embodiment, the
disintegration time is greater than 400 seconds when orally
administered. Disintegration time can be determined in water at
37.+-.2.degree. C. using the method described in USP <701>.
Disintegration times may range from about 1 second to as much as
about 24 hours, or more, depending on many factors including, but
not limited to, the particular active agent(s), delivery agent
compound(s), and excipients included in the pharmaceutical
formulation.
[0012] Another embodiment is a pharmaceutical formulation (such as
a solid oral dosage form) comprising a therapeutically effective
amount of an active agent and a delivery agent, where the solid
oral dosage form does not substantially disintegrate or dissolve in
the stomach, but does substantially disintegrate or dissolve in the
intestine. In a preferred embodiment, the active agent is insulin.
In another preferred embodiment, the active agent is an insulin
derivative.
[0013] In another embodiment, the pharmaceutical formulation is a
solid oral dosage form which is covered with an enteric coating to
retard disintegration in the stomach. Enteric coatings include, but
are not limited to, hydroxypropyl methylcellulose phthalate,
hydroxypropyl methylcellulose acetate succinate, polyvinyl acetate
phthalate, cellulose acetate trimellitate, cellulose acetate
phthalate, poly(methacrylic acid-ethylacrylate), and
poly(methacrylic acid-methyl methacrylate).
[0014] In yet another embodiment, the pharmaceutical formulations
may be formulated to erode from the surface of the dosage form,
rather than disintegrate.
[0015] The pharmaceutical formulations may include
enzyme-inhibiting agents to prevent enzymatic degradation of active
agents in the pharmaceutical formulation.
[0016] In one embodiment, the delivery agent is a compound having
the following structure or a salt thereof:
##STR00001##
[0017] wherein
[0018] Ar is phenyl or naphthyl;
[0019] Ar is optionally substituted with one or more of --OH,
halogen, C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 alkenyl,
C.sub.1-C.sub.4 alkoxy. or C.sub.1-C.sub.4 haloalkoxy;
[0020] R.sup.1 is C.sub.3-C.sub.20 alkyl, C.sub.4-C.sub.20 alkenyl,
phenyl, naphthyl, (C.sub.1-C.sub.10 alkyl) phenyl,
(C.sub.1-C.sub.10 alkenyl)phenyl, (C.sub.1-C.sub.10 alkyl)
naphthyl, (C.sub.1-C.sub.10 alkenyl) naphthyl,
phenyl(C.sub.1-C.sub.10alkyl), phenyl(C.sub.1-C.sub.10 alkenyl),
naphthyl(C.sub.1-C.sub.10 alkyl), or naphthyl(C.sub.1-C.sub.10
alkenyl);
[0021] R.sup.1 is optionally substituted with C.sub.1 to C.sub.4
alkyl, C.sub.2 to C.sub.4 alkenyl, C.sub.1 to C.sub.4 alkoxy,
C.sub.1-C.sub.4 haloalkoxy, --OH, --SH, --CO.sub.2R.sup.8, or any
combination thereof;
[0022] R.sup.2 is hydrogen, C.sub.1 to C.sub.4 alkyl, or C.sub.2 to
C.sub.4 alkenyl; and
[0023] R.sup.1 is optionally interrupted by oxygen, nitrogen,
sulfur or any combination thereof. The term "2-OH--Ar" in formula A
refers to a phenyl or naphthyl group having a hydroxyl group at the
2-position.
[0024] According to one embodiment, the compounds are not
substituted with an amino group in the position alpha to the acid
group.
[0025] Preferably, Ar is substituted with a halogen.
[0026] Preferably, R.sup.2 is hydrogen.
[0027] Preferably, R.sup.1 is unsubstituted.
[0028] Preferably, R.sup.1 is not interrupted.
[0029] Preferably, R.sup.1 is C.sub.1-10, C.sub.3-9, C.sub.3-7,
C.sub.3, C.sub.7, or C.sub.9 alkyl. According to one embodiment,
R.sup.1 is not branched.
[0030] Preferred delivery agent compounds include, but are not
limited to, N-(8-[2-hydroxybenzoyl]amino)caprylic acid (the free
acid of SNAC), N-(10-[2-hydroxybenzoyl]amino)decanoic acid (the
free acid of SNAC), 4-[(2-hydroxy-4-chloro-benzoyl)-amino]butanoic
acid (the free acid of 4-CNAB), and salts thereof, and solvates and
hydrates thereof. The salt can be, for example, a sodium salt, such
as a monosodium (i.e., SNAC, SNAD, or 4-CNAB) or disodium salt.
[0031] In another embodiment, the delivery agent is a compound
having the following structure or a salt thereof:
##STR00002##
wherein
[0032] R.sup.1, R.sup.2, R.sup.3, and R.sup.4 are independently H,
--OH, halogen, C.sub.1-C.sub.4 alkyl, C.sub.2-C.sub.4 alkenyl,
C.sub.1-C.sub.4 alkoxy, --C(O)R.sup.8, --NO.sub.2,
--NR.sup.9R.sup.10, or
--N.sup.+R.sup.9R.sup.10R.sup.11(R.sup.12).sup.-;
[0033] R.sup.5 is H, --OH, --NO.sub.2, halogen, --CF.sub.3,
--NR.sup.14R.sup.15, --N.sup.+R.sup.14R.sup.15R.sup.16
(R.sup.13).sup.-, amide, C.sub.1-C.sub.12 alkoxy, C.sub.1-C.sub.12
alkyl, C.sub.2-C.sub.12 alkenyl, carbamate, carbonate, urea, or
--C(O)R.sup.18;
[0034] R.sup.5 is optionally substituted with halogen, --OH, --SH,
or --COOH;
[0035] R.sup.5 is optionally interrupted by O, N, S, or
--C(O)--;
[0036] R.sup.6 is a C.sub.1-C.sub.12 alkylene, C.sub.2-C.sub.12
alkenylene, or arylene; R.sup.6 is optionally substituted with a
C.sub.1-C.sub.4 alkyl, C.sub.2-C.sub.4 alkenyl, C.sub.1-C.sub.4
alkoxy, --OH, --SH, halogen, --NH.sub.2, or --CO.sub.2R.sup.8;
[0037] R.sup.6 is optionally interrupted by O or N;
[0038] R.sup.7 is a bond or arylene;
[0039] R.sup.7 is optionally substituted with --OH, halogen,
--C(O)CH.sub.3, --NR.sup.10R.sup.11, or
--N.sup.+R.sup.10R.sup.11R.sup.12 (R.sup.13).sup.-;
[0040] R.sup.8 is H, C.sub.1-C.sub.4 alkyl, C.sub.2-C.sub.4
alkenyl, or --NH.sub.2;
[0041] R.sup.9, R.sup.10, R.sup.11, and R.sup.12 are independently
H or C.sub.1-C.sub.10 alkyl;
[0042] R.sup.13 is a halide, hydroxide, sulfate, tetrafluoroborate,
or phosphate;
[0043] R.sup.14, R.sup.15, and R.sup.16 are independently H,
C.sub.1-C.sub.10 alkyl, C.sub.1-C.sub.10 alkyl substituted with
--COOH, C.sub.2-C.sub.12 alkenyl, C.sub.2-C.sub.12 alkenyl
substituted with --COOH, or --C(O)R.sup.17;
[0044] R.sup.17 is --OH, C.sub.1-C.sub.10 alkyl, or
C.sub.2-C.sub.12 alkenyl; and
[0045] R.sup.18 is H, C.sub.1-C.sub.6 alkyl, --OH,
--NR.sup.14R.sup.15, or N.sup.+R.sup.14R.sup.15R.sup.16
(R.sup.13).sup.-.
[0046] In yet another embodiment, the delivery agent is a compound
having the following structure or a salt thereof:
##STR00003##
wherein
[0047] R.sup.1, R.sup.2, R.sup.3, R.sup.4 and R.sup.5 are
independently H, --CN, --OH, --OCH.sub.3, or halogen, at least one
of R.sup.1, R.sup.2, R.sup.3, R.sup.4 and R.sup.5 being --CN;
and
[0048] R.sup.6 is a C.sub.1-C.sub.12 linear or branched alkylene,
alkenylene, arylene, alkyl(arylene) or aryl(alkylene).
[0049] In yet another embodiment, the delivery agent is a compound
having the following structure or a salt thereof:
##STR00004##
wherein
[0050] each occurrence of X is hydrogen, halogen, hydroxyl, or
C.sub.1-C.sub.3 alkoxy,
[0051] R is substituted or unsubstituted C.sub.1-C.sub.3 alkylene
or substituted or unsubstituted C.sub.2-C.sub.3 alkenylene, and
[0052] n is an integer from 1 to 4.
[0053] In yet another embodiment, the delivery agent is a compound
having the following structure or a salt thereof:
##STR00005##
wherein
[0054] X is halogen, and R is substituted or unsubstituted
C.sub.1-C.sub.3 alkylene or substituted or unsubstituted
C.sub.2-C.sub.3 alkenylene.
[0055] Preferred delivery agent compounds include but are not
limited to, N-(8-[2-hydroxybenzoyl]-amino)caprylic acid,
N-(10-[2-hydroxybenzoyl]-amino)decanoic acid,
8-(2-hydroxy-4-methoxybenzoylamino)octanoic acid,
8-(2-hydroxy-5-chlorobenzoylamino)-octanoic acid,
4-[(2-hydroxy-4-chlorobenzoyl)amino]butanoic acid, and
pharmaceutically acceptable salts thereof. The pharmaceutical
formulations of the present invention may include any of the
aforementioned delivery agent compounds, or any other delivery
agent compounds, alone or in combination with one or more
additional delivery agent compounds.
[0056] Suitable active agents include but are not limited to,
proteins, polypeptides, peptides, hormones, polysaccharides, as
well as synthetic, natural or recombinant sources thereof: growth
hormones; growth hormone releasing hormones; growth hormone
releasing factor, interferons; interleukin-1; interleukin-2;
insulin, optionally having counter ions including zinc, sodium,
calcium and ammonium; insulin-like growth factor; heparin;
calcitonin; erythropoietin; atrial naturetic factor; antigens;
monoclonal antibodies; somatostatin; protease inhibitors;
adrenocorticotropin, gonadotropin releasing hormone; oxytocin;
leutinizing-hormone-releasing-hormone; follicle stimulating
hormone; glucocerebrosidase; thrombopoietin; filgrastim;
prostaglandins; cyclosporin; vasopressin; cromolyn sodium;
vancomycin; desferrioxamine; bisphosphonates; parathyroid hormone;
anti-migraine agents; glucagon-like peptide 1 (GLP-1);
antimicrobials; vitamins; and analogs, fragments, mimetics or
polyethylene glycol (PEG)-modified derivatives of these compounds;
or any combination thereof. Preferred active agents include, but
are not limited to, insulin and heparin (including, but not limited
to, unfractionated heparin and low molecular weight heparin).
[0057] In one embodiment of the present invention, the active agent
is insulin. The insulin-containing pharmacuetical formulations of
the present invention may also include a second hypoglycemic agent,
an inhibitor of renal glucose reabsorption, or any combination of
the foregoing (such as those described in U.S. Patent Publication
No. 2005/0143424, which is hereby incorporated by reference).
Suitable second hypoglycemic agents include, but are not limited
to, insulin secretion-promoting agents, insulin
resistance-ameliorating agents, insulin mimetics,
.alpha.-glucosidase inhibitors, glucogenesis inhibitors, and any
combination of any of the foregoing. According to one embodiment,
the solid dosage form includes a sulfonyl urea, meglitinide
analogue, biguanide (preferably metformin), or any combination of
any of the foregoing. According to a preferred embodiment, the
solid dosage form includes metformin.
[0058] Also provided is a pharmaceutical formulation, such as a
solid dosage unit form, comprising the microparticles or
nanoparticles of the present invention and/or having the
disintegration times discussed above. The dosage unit form may be
in the form of a tablet, capsule, powder, or sachet. The dosage
unit form may have, alone or in combination, one or more enteric
coatings, disintegrants, super disintegrants (such as sodium starch
glycolate or croscarmellose sodium), and extra particle super
disintegrants.
[0059] In one embodiment, the solid oral dosage unit form is a fast
disintegrating tablet. In another embodiment, the solid dosage unit
form has a controlled or delayed release.
[0060] According to one embodiment, the present invention provides
a tablet comprising the aforementioned particles and a
disintegrant. In one embodiment, the disintegrant is a super
disintegrant, such as sodium starch glycolate (Primojel.RTM.
available from Azebe UK Ltd. of South Humberside, UK),
croscarmellose sodium (Primellose.RTM. available from Azebe UK Ltd.
of South Humberside, UK), or an extra particle super
disintegrant.
[0061] Another embodiment is a solid dosage form comprising a
therapeutically effective amount of insulin and a delivery agent
compound, where the solid dosage form has a disintegration time of
at least 60 seconds when administered orally. The solid dosage form
may have an enteric coating or be a surface eroding formulation.
The solid dosage form may further comprise one or more enzyme
inhibiting agents.
[0062] Yet another embodiment is a solid dosage form comprising a
therapeutically effective amount of insulin and a delivery agent
compound, where the solid dosage form does not substantially
disintegrate or dissolve in the stomach but does disintegrate or
dissolve in the small intestine. The solid dosage form may have an
enteric coating or be a surface eroding formulation. The solid
dosage form may further comprise one or more enzyme inhibiting
agents.
[0063] Another embodiment is a method for administering an active
agent to an animal, particularly an animal in need of the active
agent, by administering a pharmaceutical formulation comprising the
microparticles or nanoparticles of the present invention and/or
those having the disintegration times discussed above (i.e. those
having a controlled or sustained release). Oral administration is a
preferred route of administration.
[0064] Yet another embodiment is a method of treating a disease or
for achieving a desired physiological effect in an animal by
administering a pharmaceutical formulation of the present
invention, including solid unit dosage forms comprising the
microparticles or nanoparticles of the present invention and/or
those having the disintegration times discussed above (i.e. those
having a controlled or sustained release). Yet another embodiment
is a method of increasing the oral bioavailability of active agents
by orally administering a pharmaceutical formulation of the present
invention.
[0065] Yet another embodiment is a method of treating diabetes
and/or reducing the incidence of systemic hyperinsulinemia
associated with chronic dosing of insulin in a mammal (such as in a
human, particularly a human in need thereof) by administering to
the mammal a therapeutic effective amount of an insulin-containing
pharmaceutical formulation of the present invention, e.g., those
comprising the microparticles or nanoparticles of the present
invention and/or those having the disintegration times discussed
above. In one embodiment, the delivery agent compound is the free
acid of 4-CNAB or a pharmaceutically acceptable salt thereof. The
pharmaceutical formulation may be administered on a chronic
basis.
[0066] Yet another embodiment is a method of treating impaired
glucose tolerance, early stage diabetes, or late stage diabetes or
achieving glucose homeostasis in a mammal (such as in a human,
particularly in need thereof) by administering to the mammal a
therapeutic effective amount of an insulin-containing
pharmaceutical formulation of the present invention, such as a
pharmaceutical formulation comprising the microparticles or
nanoparticles of the present invention and/or having the
disintegration times discussed above. In one embodiment, the
delivery agent compound is the free acid of 4-CNAB or a
pharmaceutically acceptable salt thereof. The pharmaceutical
formulation may be administered on a chronic basis.
[0067] Yet another embodiment is a method of treating a human
diabetic patient by orally administering to the human diabetic
patient on a chronic basis a therapeutic effective amount of an
insulin-containing pharmaceutical formulation described herein.
[0068] Yet another embodiment is a method of preparing the micro-
and nano-particles of the present invention by drying a solution of
a delivery agent compound and an active agent, for example, until a
solid is formed, and optionally, isolating the particles.
Preferably, the mixture is homogenous (e.g., the delivery agent
compound and the active agent are uniformly distributed throughout
the mixture). The method includes co-drying a mixture of the
delivery agent compound, the active agent, and a solvent. Suitable
solvents include, but are not limited to, hydroxylic solvents,
water, and mixtures thereof. According to one embodiment, the
mixture is dried at from about 10 to about 40.degree. C. (e.g., at
room temperature). Preferably, the drying is performed at a
controlled temperature. According to one embodiment, the drying is
performed over an inert gas (preferably nitrogen gas). The dried
material may optionally be milled and/or sieved to obtain the
desired particle size. This method results in particles containing
a homogeneous mixture of the delivery agent compound and the active
agent.
[0069] Another method of preparing the micro- and nano-particles of
the present invention is by lyophilizing a mixture of the delivery
agent compound, the active agent, and a solvent. Suitable solvents
include, but are not limited to, hydroxylic solvents, water, and
mixtures thereof.
[0070] Yet another method of preparing the micro- and
nano-particles of the present invention is by (1) dissolving a
delivery agent compound and an active agent in a supercritical
fluid, and (2) decreasing the system pressure to deposit the
delivery agent compound and active agent as extremely fine
particles. The deposition is a result of the rapid expansion of the
supercritical solution.
[0071] The following embodiments are collectively referred to
herein as the "solid pharmaceutical composition embodiments".
[0072] Yet another embodiment is a solid pharmaceutical composition
which enhances the oral bioavailability of active agents,
particularly peptides. More specifically, the solid pharmaceutical
composition suitable for the oral delivery of pharmacologically
active agents, comprises:
[0073] 1. a therapeutically-effective amount of a pharmacologically
active agent;
[0074] 2. one or more pharmaceutically acceptable inactive
excipients; and
[0075] 3. a delivery agent for the pharmacologically active agent,
wherein the delivery agent is in micronized form.
[0076] Yet another embodiment is a solid pharmaceutical composition
suitable for the oral delivery of calcitonin, comprising:
[0077] 1. a therapeutical-effective amount of a calcitonin; and
[0078] 2. one or more pharmaceutically acceptable inactive
excipients, and
[0079] 3. a delivery agent for said calcitonin, wherein said
delivery agent is in micronized form.
[0080] In an additional embodiment, the pharmaceutically acceptable
inactive excipient may be either or both of the polymers
crospovidone or povidone.
[0081] In a still further embodiment, the solid pharmaceutical
composition suitable for oral delivery may also comprise a
diluent.
[0082] In addition, in another embodiment the solid pharmaceutical
composition suitable for oral delivery may also comprise a
lubricant.
[0083] In a further embodiment, the invention is directed to a
method for enhancing the oral bioavailability of a
pharmacologically active agent. The method comprises administering
to a subject in need of the pharmacologically active agent an
effective amount of a pharmaceutical composition according to the
instant invention.
[0084] Yet another embodiment is a method of treatment of bone
related diseases and calcium disorders comprising administering to
a patient in need of such treatment a therapeutically effective
amount of a composition according to the instant invention, wherein
the pharmacologically active agent is calcitonin.
[0085] The above features and many other attendant advantages of
the invention will become better understood by reference to the
following detailed description when taken in conjunction with the
accompanying drawings.
BRIEF DESCRIPTION OF THE DRAWINGS
[0086] FIG. 1 depicts a schematic of direct dosing to the stomach
and the jejunum.
[0087] FIG. 2 is a graph of the concentration of insulin level
(.+-.SEM) following direct dosing of coprocessed microparticles to
the stomach and the jejunum over time.
[0088] FIG. 3 is a graph of the change in glucose level (.+-.SEM)
following direct dosing of coprocessed microparticles to the
stomach and the jejunum over time.
[0089] FIG. 4 is a graph of the change in glucose (.+-.SEM)
following oral gavage from 3 different dosage forms: 1) a tablet
made by compressing insulin and carrier, 2) a capsule containing
microparticles of coprocessed insulin and carrier, and 3) a capsule
containing a simple mixture of insulin and carrier, over time.
[0090] FIG. 5 is a graph of the insulin level (.+-.SEM) following
oral gavage from 3 different dosage forms: 1) a tablet made by
compressing insulin and carrier, 2) a capsule containing
microparticles of coprocessed insulin and carrier, and 3) a capsule
containing a simple mixture of insulin and carrier, over time.
[0091] FIG. 6 is a graph of the insulin level (ASEM) following oral
gavage of a capsule containing microparticles of coprocessed
insulin and carrier over time. Two of the ten rats exhibited
significantly high insulin absorption. The average values with
(N=10) and without (N=8) inclusion of these two high responders are
depicted in the graph.
[0092] FIG. 7 is a graph of the change in glucose (.+-.SEM)
following oral gavage of a capsule containing microparticles of
coprocessed insulin and carrier over time. Two of the ten rats
exhibited significantly high insulin absorption. The average values
with (N=10) and without (N=8) inclusion of these two high
responders are depicted in the graph.
[0093] FIG. 8 is a chart of the estimated absolute bioavailability
(.+-.SEM) from in situ dosing of coprocessed insulin and carrier to
the stomach and the jejunum. Two compositions were evaluated: 1)
insulin (0.25 mg/kg)+delivery agent (37.5 mg/kg), and 2) insulin
(0.5 mg/kg)+delivery agent (75 mg/kg).
[0094] FIG. 9 is a chart of the estimated absolute bioavailability
of insulin level (.+-.SEM) from 1) subcutaneous administration, 2)
direct dosing to the stomach, 3) direct dosing to the jejunum, 4) a
tablet made by compressing insulin and carrier, 5) a capsule
containing microparticles of coprocessed insulin and carrier with
and without inclusion of the two high responders, and 6) a capsule
containing a simple mixture of insulin and carrier.
[0095] FIG. 10 is a chart of the estimated bioavailability of
insulin in the portal vein (.+-.SEM) from 1) direct dosing to the
stomach, 2) direct dosing to the jejunum, 3) a tablet made by
compressing insulin and carrier, 4) a capsule containing
microparticles of coprocessed insulin and carrier with and without
inclusion of the two high responders, and 5) a capsule containing a
simple mixture of insulin and carrier.
[0096] FIG. 11 is a chart of the estimated bioavailability
(.+-.SEM) of insulin relative to subcutaneous administration from
1) direct dosing to the stomach, 2) direct dosing to the jejunum,
3) a tablet made by compressing insulin and carrier, 4) a capsule
containing microparticles of coprocessed insulin and carrier with
and without inclusion of the two high responders, and 5) a capsule
containing a simple mixture of insulin and carrier.
[0097] FIG. 12 is a chart of the estimated bioavailability of
insulin in the portal vein relative to subcutaneous administration
(.+-.SEM) from 1) direct dosing to the stomach, 2) direct dosing to
the jejunum, 3) a tablet made by compressing insulin and carrier,
4) a capsule containing microparticles of coprocessed insulin and
carrier with and without inclusion of the two high responders, and
5) a capsule containing a simple mixture of insulin and
carrier.
[0098] FIG. 13 is a graph of the individual insulin levels
following oral gavage of a capsule containing microparticles of
coprocessed insulin and carrier over time. Rat 14 and rat 17
exhibited significantly high insulin absorption. The average values
with (N=10) and without (N=8) inclusion of these two high
responders are depicted in the graph.
[0099] FIG. 14 is a graph of the individual glucose change
following oral gavage of a capsule containing microparticles of
coprocessed insulin and carrier over time. Rat 14 and rat 17
exhibited significantly high insulin absorption. The average values
with (N=10) and without (N=8) inclusion of these two high
responders are depicted in the graph.
[0100] FIG. 15 is a graph of the individual insulin level following
oral gavage of a capsule containing microparticles of coprocessed
insulin and carrier over time. Rats 14 and 17 were omitted. The
average value from N=8 is depicted in the graph.
[0101] FIG. 16 is a graph of the individual glucose change
following oral gavage of a capsule containing microparticles of
coprocessed insulin and carrier over time. Rats 14 and 17 were
omitted. The average value from N=8 is depicted in the graph.
[0102] FIG. 17 is a graph depicting the changes over time in serum
glucose levels in rhesus monkeys that have been fed formulations
1-6, described below, containing insulin and a delivery agent.
These formulations have varying disintegration times.
[0103] FIG. 18 is a graph depicting the changes over time in serum
insulin concentration rhesus monkeys that have been fed
formulations 1-6, described below, containing insulin and a
delivery agent. These formulations have varying disintegration
times.
[0104] FIG. 19 is a graph of anti-factor Xa activity (U/ml) versus
time in monkeys after administration of the SNAD/heparin
formulation described in Example 10.
DETAILED DESCRIPTION OF THE INVENTION
Definitions
[0105] The "particles," "micro-beads," and "granules" described
herein may be any shape and can include one or more ingredients in
addition to the delivery agent compound and/or active agent. The
specific ingredients of any given particle, micro-bead, or granule,
may also depend on the processes used and will not necessarily be
the same in each individual particle, micro-bead, or granule from a
batch.
[0106] For example, where particles, micro-beads, or granules of an
active agent are prepared separately from particles, micro-beads,
or granules of a delivery agent compound, the active agent
particles, micro-beads, or granules will, generally, not comprise
delivery agent compound, and the delivery agent particles,
micro-beads, or granules will, generally, not comprise active
agent, though each particle, micro-bead, or granule may comprise
other ingredients, as disclosed herein.
[0107] In other embodiments, particles, micro-beads, or granules
may be formed from a solution, suspension or mixture, in liquid or
dry form, without limitation, which comprises at least an active
agent and a delivery agent compound. Thus, for example, any given
particle, micro-bead, or granule comprises both active agent and
delivery agent compound, and may further comprise one or more other
ingredients.
[0108] The terms "diameter" and "median particle size" are
generally used to refer to the dimensions of particles,
micro-beads, and granules. The "median particle size" or "diameter"
was determined as follows for Examples 8, 9, 10.
[0109] Instrument: Mastersizer 2000 (EQ 202, model MS2K, serial
number 34315-67)
[0110] Manufacturer: MALVERN instruments, England
[0111] Software: Mastersizer 2000
[0112] Accessory: Scirocco 2000 (A) (model ADA 2000, serial number
34270/73)
[0113] Dispersant: Dry dispersion
[0114] Analysis model: General purpose
[0115] Particle RI: 1.520
[0116] Obscuration: 1-6%
[0117] Standards: Malvem Quality Audit Standard for Sample
Dispersion Units
[0118] The Malvern Mastersizer 2000 determines particle size by
laser diffraction and model fitting. A well-dispersed sample in any
two-phase system (e.g., powders, suspensions, or emulsions) is
introduced into the path of a He--Ne laser focused with a lens of a
length suitable for particle sizes present in the sample. The
scattering pattern of particles in the laser path is measured by an
array of detectors, with each detector measuring data from a
particular range of angles.
[0119] The Malvern apparatus assumes that the particles being
measured are perfect spheres. For non-spherical particles the
resulting particle size distribution may be different from those
obtained by methods based on others principles. The electronic
measurements will often have to be accompanied by microscopic
investigation to determine the type of particles being
investigated. For irregularly shaped particles, the particle size
data obtained from Mastersizer 2000 will be interpreted as the
diameter of an imaginary sphere that is equivalent in volume to the
measured particle. (Note: d(0.1) is the size of particle for which
10% of the sample is below this size, d(0.5) is the size of
particle for which 50% of the sample is below this size, and d(0.9)
is the size of particle for which 90% of the sample is below this
size.
[0120] Generally, this apparatus measures one dimension of a, e.g.,
particle as it travels past a laser; i.e., it measures the length
of a straight line through the particle. For irregular particles,
this results in a variation of results since the orientation of a
particle relative to the laser may result in the single measurement
being taken of that individual particle's longest, shortest, or any
other dimension. However, a measurement is taken of a number of
particles and a median diameter or size is calculated. Thus, "size"
or "diameter" figures are estimates of the median "size" or
"diameter" of particles. Alternatively, "diameter" or "size" was
measured by a sieve method described in Example 1. "Diameter"
should not be read to necessarily imply a spherical shape or a
circular dimension, though in certain embodiments, e.g., particles
may have rounded edges or generally spherical shapes.
[0121] It should be understood, also, that the invention is not
limited to particles, micro-beads, or granules which fall within a
narrow range of "sizes" or "diameters". Thus, for example, some
embodiments may comprise, depending at least on the ingredients and
processes used, some particles which fall within, for example, both
the nanometer and micrometer scale, in the same batch. The actual
"sizes" or "diameters" of the individual particles may fall within
a relatively narrow or relatively large range.
[0122] As used herein and in the appended claims, the singular
forms "a," "an," and "the," include plural referents unless the
context clearly indicates otherwise. Thus, for example, reference
to "a particle" includes one or more of such particles, reference
to "an" active agent includes one or more of such active agents,
and "a" delivery agent includes one or more delivery agents,
[0123] The term "about" generally means within 10%, preferably
within 5%, and more preferably within 1% of a given value or
range.
[0124] The term "hydrate" as used herein includes, but is not
limited to, (i) a substance containing water combined in the
molecular form and (ii) a crystalline substance containing one or
more molecules of water of crystallization or a crystalline
material containing free water.
[0125] The term "solvate" as used herein includes, but is not
limited to, a molecular or ionic complex of molecules or ions of a
solvent with molecules or ions of the delivery agent compound or
salt thereof, or hydrate or solvate thereof.
[0126] The term "delivery agent" refers to any of the delivery
agent compounds disclosed herein.
[0127] The term "SNAC" refers to the monosodium salt of
N-(8-[2-hydroxybenzoyl]-amino)caprylic acid, including the various
polymorphic forms of the monosodium salt described in U.S.
Provisional Application No. 60/569,476, filed May 6, 2004 (which is
hereby incorporated by reference) unless otherwise indicated.
[0128] The term "SNAD" refers to the monosodium salt of
N-(10-[2-hydroxybenzoyl]-amino)decanoic acid, unless otherwise
indicated. The term "disodium salt of SNAD" refers to the disodium
salt of N-(10-[2-hydroxybenzoyl]-amino)decanoic acid.
[0129] The term "5-CNAC" refers to the monosodium salt of
N-(8-[2-hydroxy-5-chlorobenzoyl]-amino)octanoic acid, unless
otherwise indicated.
[0130] The term "4-CNAB" refers to the monosodium salt of sodium
N-4-[(2-hydroxy-4-chlorobenzoyl)amino]butanoate, including
anhydrous, monohydrate, and isopropanol solvates thereof and
various polymorphic forms of the monosodium salt described in
International Publication No. WO 03/057650 (which is hereby
incorporated by reference), unless otherwise indicated.
[0131] An "effective amount of active agent" is an amount of active
agent which is effective to treat or prevent a condition in a
living organism to whom it is administered over some period of
time, e.g., provides a therapeutic effect during a desired dosing
interval.
[0132] The term "insulin" refers to all forms of insulin,
including, but not limited to, naturally derived insulin and
synthetic forms of insulin, such as those described in U.S. Pat.
Nos. 4,421,685, 5,474,978, and 5,534,488, each of which is hereby
incorporated by reference in its entirety.
[0133] The term "insulin derivatives" refers to insulin-derived
proteins and peptides with insulin actions, and include, for
example, lispro, B10Asp and HOE-901.
[0134] An "effective amount of delivery agent" is an amount of the
delivery agent which enables and/or facilitates the absorption of a
desired amount of active agent via any route of administration
(such as those discussed in this application including, but not
limited to, the oral (e.g., across a biological membrane in the
gastrointestinal tract), nasal, pulmonary, dermal, buccal, vaginal,
and/or ocular route).
[0135] The terms "alkyl" and "alkenyl" as used herein include
linear and branched alkyl and alkenyl substituents,
respectively.
[0136] The phrase "pharmaceutically acceptable" refers to additives
or compositions that are physiologically tolerable when
administered to a mammal.
[0137] The phrase "substantially disintegrate" means that about 75%
to about 95% of the total volume of the tablet will break apart and
dissolve into its component parts (e.g. insoluble coated particles,
insoluble disintegrant, etc.), and the tablet is no longer intact
except for small aggregates.
[0138] "Surface eroding formulation" refers to formulations that do
not disintegrate but instead erode, e.g., the formulation dissolves
from the surface over a pre-determined period of time and the
tablet generally remains intact and retains its overall shape. The
surface eroding formulations allow for sustained release of an
active agent over the pre-determined time period.
[0139] The terms "micronize" and "micronized" generally refer to a
process, or particles which have been processed, such that their
diameters/sizes are within the general range of microparticles
and/or nanoparticles.
[0140] The term "microparticle" generally includes particles having
a diameter ranging from about 1 to about 999 micrometers (microns,
.mu.m).
[0141] The term "nanoparticle" generally includes particles having
a diameter ranging from about 1 to about 999 nanometers (nm).
[0142] The term "insulin derivatives" includes insulin-derived
proteins and peptides with insulin actions, and include, for
example, lispro, B10Asp and HOE-901.
[0143] "Insulin secretion-promoting agents" exert their
hypoglycemic action, by mainly influencing pancreatic n-cells to
promote insulin secretion into blood, and include, for example,
sulfonylureas (for example, tolbutamide, chlorpropamide,
glibenclamide (glyburide), glipizide, glimeperide, and gliclazide);
and meglitinide analogues (for example, repaglinide, nateglinide,
meglitinide and mitiglinide (KAD-1229))). Other insulin
secretion-promoting agents are, for example, K+-ATP channel
inhibitors (for example, BTS-67-582), glucagon-like peptide-1
receptor agonists (for example, glucagon-like peptide-1, exendin-4
and NN-2211) and dipeptidyl peptidase-IV inhibitors with an effect
of enhancing the action of glucagon-like peptide-1. According one
embodiment, the insulin secretion-promoting agent is a sulfonylurea
or meglitinide analogue.
[0144] The term "insulin resistance-ameliorating agents" includes
agents exerting hypoglycemic action by enhancing the action of
insulin in target tissues, and include for example peroxisome
proliferator activator receptor (PPAR)-.gamma. agonists (for
example, thiazolidine-based compounds such as pioglitazone,
rosiglitazone, and ciglitazone; or non-thiazolidine-based compounds
such as GI-262570, JTT-501, YM-440, NN-622 and KRP-297),
PPAR-.gamma. antagonists and protein tyrosine phosphatase
inhibitors. The insulin resistance-ameliorating agents include, for
example, pharmaceutical agents with a function ameliorating insulin
resistance, for example biguanides (for example, metformin,
phenformin and buformin, preferably metformin), PPAR-.alpha.
agonists (fibrate-series compounds such as simfibrate, clofibrate,
bezafibrate and clinofibrate and non-fibrate-series compounds),
anti-obesity agents (for example, 5-hydroxytryptamine (5-HT)
reuptake inhibitors such as sibutramine, lipase inhibitors such as
orlistat and adrenalin .beta.-receptor agonists such as AJ-9677).
Preferred insulin resistance-ameliorating agents include, but are
not limited to, biguanides, such as metformin.
[0145] The term "insulin mimetics" refers to agents expressing the
hypoglycemic action through physiological insulin action, namely
the action promoting glucose uptake into cells, in a manner more or
less independent to insulin, except for insulin derivatives, and
include for example insulin receptor-activating agents (for
example, CLX-0901 and L-783281) and vanadium.
[0146] The term ".alpha.-glucosidase inhibitors" refers to agents
expressing the hypoglycemic action through suppression of glucose
absorption into bodies, mainly via the inhibition of
.alpha.-glucosidase in the intestinal tube and include, for
example, acarbose, voglibose and miglitol.
[0147] The term "glucogenesis inhibitors" refers to agents
expressing hypoglycemic action mainly through the inhibition of
glucogenesis, and include for example glucagon secretion
suppressors (for example, M&B-39890A and octreotide), fatty
acid decomposition inhibitors (for example, nicotinic acid
derivatives and camitine palmitoyltransferase-1 inhibitor) and
glucose-6-phosphatase inhibitors.
[0148] The term "inhibitor of renal glucose reabsorption" refers to
agents which inhibit glucose reabsorption in uriniferous tubules.
The primary action of the inhibitor of renal glucose reabsorption
is not involved in the promotion of the uptake into target tissue
cells, the suppression of the absorption from intestinal tube, or
the hypoglycemic action via the suppression of the synthesis in
tissues. Suitable inhibitors of renal glucose reabsorption include,
but are not limited to, those described in U.S. Patent Publication
No. 2005/0143424, which is hereby incorporated by reference.
Delivery Agent Compounds
[0149] The delivery agent compound may be any of those described in
U.S. Pat. Nos. 5,650,386 and 5,866,536 and International
Publication Nos. WO94/23767, WO95/11690, WO95/28920, WO95/28838,
WO96/10396, WO96/09813, WO96/12473, WO96/12475, WO96/30036,
WO96/33699, WO97/31938, WO97/36480, WO98/21951, WO98/25589,
WO98/34632, WO98/49135, WO99/16427, WO00/06534, WO00/07979,
WO00/40203, WO00/46182, WO00/47188, WO00/48589, WO00/50386,
WO00/59863, WO00/59480, WO01/32130, WO01/32596, WO01/34114,
WO01/44199, WO01/51454, WO01/70219, WO01/92206, WO02/02509,
WO02/15959, WO02/16309, WO02/20466, WO02/19969, WO02/070438,
WO03/026582, WO02/100338, WO03/045306, WO03/26582, and WO
03/057170, all of which are hereby incorporated by reference.
[0150] Non-limiting examples of delivery agent compounds include
N-(8-[2-hydroxybenzoyl]amino)caprylic acid,
N-(10-[2-hydroxybenzoyl]amino)decanoic acid,
8-(2-hydroxy-4-methoxybenzoylamino)octanoic acid,
8-(2-hydroxy-5-chlorobenzoyl-amino)octanoic acid,
4-[(2-hydroxy-4-chlorobenzoyl)amino]butanoic acid, and salts
thereof. Preferred salts include, but are not limited to,
monosodium and disodium salts.
[0151] According to one embodiment, the delivery agent compound is
N-(8-[2-hydroxybenzoyl]amino)caprylic acid or a pharmaceutically
acceptable salt thereof.
[0152] According to another embodiment, the delivery agent compound
is N-(10-[2-hydroxybenzoyl]amino)decanoic acid or a
pharmaceutically acceptable salt thereof.
[0153] According to another embodiment, the delivery agent compound
is 4-[(2-hydroxy-4-chlorobenzoyl)amino]butanoic acid or a
pharmaceutically acceptable salt thereof.
[0154] According to another embodiment, the delivery agent compound
is 8-(2-hydroxy-5-chlorobenzoylamino)octanoic acid or a
pharmaceutically acceptable salt thereof.
[0155] The delivery agent compounds may be in the form of the
carboxylic acid or pharmaceutically acceptable salts thereof, such
as sodium salts, and hydrates and solvates thereof. The salts may
be mono- or multi-valent salts, such as monosodium salts and
disodium salts (e.g., the disodium salt of
8-(2-hydroxy-5-chlorobenzoylamino)-octanoic acid, the disodium salt
of N-(8-[2-hydroxybenzoyl]amino)caprylic acid, the disodium salt of
N-(10-[2-hydroxybenzoyl]amino)decanoic acid). See, for example,
International Publication No. WO 00/59863, which is hereby
incorporated by reference The delivery agent compounds may contain
different counter ions chosen for example due to their effect on
modifying the dissolution profile of the carrier.
[0156] The delivery agent compounds may be prepared by methods
known in the art, such as those discussed in the aforementioned
publications (e.g., International Publication Nos. WO 98/34632, WO
00/07979, WO 01/44199, WO 01/32596, WO 02/02509, WO 02/20466, and
WO 03/045306). SNAC, SNAD, 4-CNAB, and the free acid and other
salts thereof may be prepared by methods known in the art, such as
those described in U.S. Pat. Nos. 5,650,386 and 5,866,536 and
International Publication No. WO 02/02509, each of which are hereby
incorporated by reference.
[0157] Salts of the delivery agent compounds of the present
invention may be prepared by methods known in the art. For example,
sodium salts may be prepared by dissolving the delivery agent
compound in ethanol and adding aqueous sodium hydroxide.
[0158] The delivery agent compound may be purified by
recrystallization or by fractionation on one or more solid
chromatographic supports, alone or linked in tandem. Suitable
recrystallization solvent systems include, but are not limited to,
acetonitrile, methanol, and tetrahydrofuran. Fractionation may be
performed on a suitable chromatographic support such as alumina,
using methanol/n-propanol mixtures as the mobile phase; reverse
phase chromatography using trifluoroacetic acid/acetonitrile
mixtures as the mobile phase; and ion exchange chromatography using
water or an appropriate buffer as the mobile phase. When anion
exchange chromatography is performed, preferably a 0-500 mM sodium
chloride gradient is employed.
[0159] The delivery agent may contain a polymer conjugated to it by
a linkage group selected from the group consisting of --NHC(O)NH--,
--C(O)NH--, --NHC(O), --OOC--, --COO--, --NHC(O)O--, --OC(O)NH--,
--CH.sub.2NH--NHCH.sub.2--, --CH.sub.2NHC(O)O--,
--OC(O)NHCH.sub.2--, --CH.sub.2NHCOCH.sub.2O--,
--OCH.sub.2C(O)NHCH.sub.2--, --NHC(O)CH.sub.2O--,
--OCH.sub.2C(O)NH--, --NH--, --O--, and carbon-carbon bond, with
the proviso that the polymeric delivery agent is not a polypeptide
or polyamino acid. The polymer may be any polymer including, but
not limited to, alternating copolymers, block copolymers and random
copolymers, which are safe for use in mammals. Preferred polymers
include, but are not limited to, polyethylene; polyacrylates;
polymethacrylates; poly(oxyethylene); poly(propylene);
polypropylene glycol; polyethylene glycol (PEG); and derivatives
thereof and combinations thereof. The molecular weight of the
polymer typically ranges from about 100 to about 200,000 daltons.
The molecular weight of the polymer preferably ranges from about
200 to about 10,000 daltons. In one embodiment, the molecular
weight of the polymer ranges from about 200 to about 600 daltons
and more preferably ranges from about 300 to about 550 daltons.
Active Agents
[0160] Active agents suitable for use in the present invention
include biologically active agents and chemically active agents,
including, but not limited to, pesticides, pharmacological agents,
and therapeutic agents. Suitable active agents include those that
are rendered less effective, ineffective or are destroyed in the
gastro-intestinal tract by acid hydrolysis, enzymes and the like.
Also included as suitable active agents are those macromolecular
agents whose physiochemical characteristics, such as, size,
structure or charge, prohibit or impede absorption when dosed
orally.
[0161] For example, biologically or chemically active agents
suitable for use in the present invention include, but are not
limited to, proteins; polypeptides; peptides; hormones;
polysaccharides, and particularly mixtures of muco-polysaccharides;
carbohydrates; lipids; small polar organic molecules (i.e. polar
organic molecules having a molecular weight of 500 daltons or
less); other organic compounds; and particularly compounds which by
themselves do not pass (or which pass only a fraction of the
administered dose) through the gastro-intestinal mucosa and/or are
susceptible to chemical cleavage by acids and enzymes in the
gastro-intestinal tract; or any combination thereof.
[0162] Further examples include, but are not limited to, the
following, including synthetic, natural or recombinant sources
thereof: growth hormones, including human growth hormones (hGH),
recombinant human growth hormones (rhGH), bovine growth hormones,
and porcine growth hormones; growth hormone releasing hormones;
growth hormone releasing factor, interferons, including .alpha.
(e.g., interferon alfacon-1 (available as Infergen.RTM. from
InterMune, Inc. of Brisbane, Calif.)), .beta. and .gamma.;
interleukin-1; interleukin-2; insulin, including porcine, bovine,
human, and human recombinant, optionally having counter ions
including zinc, sodium, calcium and ammonium; insulin-like growth
factor, including IGF-1; heparin, including unfractionated heparin,
heparinoids, dermatans, chondroitins, low molecular weight heparin,
very low molecular weight heparin and ultra low molecular weight
heparin; calcitonin, including salmon, eel, porcine and human;
erythropoietin; atrial naturetic factor; antigens; monoclonal
antibodies; somatostatin; protease inhibitors; adrenocorticotropin,
gonadotropin releasing hormone; oxytocin;
leutinizing-hormone-releasing-hormone; follicle stimulating
hormone; glucocerebrosidase; thrombopoietin; filgrastim;
prostaglandins; cyclosporin; vasopressin; cromolyn sodium (sodium
or disodium chromoglycate); vancomycin; desferrioxamine (DFO);
bisphosphonates, including alendronate, tiludronate, etidronate,
clodronate, pamidronate, olpadronate, and incadronate; parathyroid
hormone (PTH), including its fragments; anti-migraine agents such
as BIBN-4096BS and other calcitonin gene-related proteins
antagonists; glucagon-like peptide 1 (GLP-1); antimicrobials,
including antibiotics, anti-bacterials and anti-fungal agents;
vitamins; analogs, fragments, mimetics or polyethylene glycol
(PEG)-modified derivatives of these compounds; or any combination
thereof. Non-limiting examples of antibiotics include gram-positive
acting, bacteriocidal, lipopeptidal and cyclic peptidal
antibiotics, such as daptomycin and analogs thereof.
[0163] According to one embodiment, the active agent is
insulin.
[0164] According to another embodiment, the active agent is
heparin, such as unfractionated heparin or low molecular weight
heparin.
[0165] The amount of active agent used in a pharmaceutical
composition or dosage unit form of the present invention is an
amount effective to treat the target indication. However, the
amount can be less than that amount when the composition is used in
a dosage unit form because the dosage unit form may contain a
plurality of delivery agent compound/active agent, such
compositions may contain a divided effective amount. The total
effective amount can then be administered in cumulative units
containing, in total, an effective amount of active agent.
Moreover, those skilled in the field will recognize that an
effective amount of active agent will vary with many factors
including the age and weight of the animal, the animal's physical
condition, as well as other factors.
[0166] The total amount of active agent to be used of can be
determined by methods known to those skilled in the art. However,
because the compositions of the invention may deliver active agent
more efficiently than compositions containing the active agent
without the delivery agent, lower amounts of active agent than
those used in prior dosage unit forms or delivery systems can be
administered to the subject, while still achieving the same blood
levels and/or therapeutic effects.
[0167] According to one embodiment, insulin is administered at a
dose of about 0.025 to about 1.0 mg per kilogram of body weight of
the recipient per day (mg/kg/day), about 0.06 to about 0.25
mg/kg/day, or about 0.09 to about 0.19 mg/kg/day (based on the
weight of active agent). The desired dose may be administered
either as a single or divided dose.
[0168] Generally an effective amount of delivery agent to
facilitate the delivery of the active agent is administered with
the active agent. According to one embodiment, the amount of
delivery agent to active agent on a molar basis ranges from about
100:1 to about 1:1, from about 80:1 to about 2:1, or from about
20:1 to about 10:1. Delivery agent to active agent molar basis
ranges may be higher than 100:1 for particular combinations of
delivery agents and active agents. Alternatively, delivery agent to
active agent ranges may be about 1:1 or lower, such as, e.g., 0.1:1
or lower, with particular combinations of delivery agents and
active agents.
[0169] Dosage unit forms can also include any one or combination of
excipients, disintegrants, lubricants, plasticizers, colorants,
flavorants, taste-masking agents, sugars, sweeteners, and
salts.
[0170] The compositions of the subject invention are useful for
administering biologically or chemically active agents to any
animals, including but not limited to birds such as chickens,
insects, fish, reptiles, mammals (including, but not limited to,
rodents, aquatic mammals, domestic animals such as dogs and cats,
farm animals such as sheep, pigs, cows and horses, and preferably
humans).
[0171] Another embodiment of the present invention is a method for
the treatment or prevention of a disease or for achieving a desired
physiological effect, such as those listed in the table 1 below, in
an animal by administering the particles of the present invention.
Preferably, an effective amount of the particles for the treatment
or prevention of the desired disease or for achieving the desired
physiological effect is administered. Specific indications for
active agents can be found in the Physicians' Desk Reference
(58.sup.th Ed., 2004, Medical Economics Company, Inc., Montvale,
N.J.), which is herein incorporated by reference. The active agents
in the table below include their analogs, fragments, mimetics, and
polyethylene glycol-modified derivatives.
TABLE-US-00001 TABLE 1 Non-Limiting Examples of Disease And Active
Agent Physiological Effect Amylin and Amylin Agonists; Obesity
Adrenocorticotropin; High Cholesterol (To Lower Cholesterol)
Antigens; Infection Antimicrobials, including Antibiotics,
Infection Including Gram-Positive Bacterial Anti-Bacterials and
Anti-Fungal Agents; Infection non-limiting examples of Antibiotics
include Gram-Positive Acting, Bacteriocidal, Lipopeptidal and
Cyclic Peptidal Antibiotics, such as Daptomycin and Analogs
thereof; Anti-Migraine Agents such as BIBN- Migraines 4096BS and
Other Calcitonin Gene- Related Proteins Antagonists, Sumatriptan
Succinate; Antivirals including Acyclovir and Viral Infections
Valacyclovir; Atrial Naturetic Factor; Vasodilation Argatroban;
Prophylaxis and treatment of thrombosis in patients with
herapin-induced throbocytopenia ("HIT"), as well as an
anticoagulant therapy in patients who have or are at risk for HIT
undergoing percutaneous coronary intervention ("PCI"). Argatroban
is also useful to treat thrombotic and isechemic stroke.
Bisphosphonates, including Alendronate, Osteoporosis; Paget's
disease; Inhibits Clodronate, Etidronate, Ibandronate, osteoclasts
and Promotes osteoblastic Incadronate, Minodronate, Neridronate,
activity; treat and/or prevent bone mineral Olpadronate,
Pamidronate, Risedronate, density (bmd) loss; Breast cancer,
including Tiludronate, Zoledronate, EB1053, and as adjuvant therapy
for early stage breast YH529; cancer; Prostate cancer,; Testicular
cancer; Colon cancer; Pancreatic cancer; Endometrial cancer; Small
cell and non- small cell cancer of the lung; Ovarian cancer;
Cervical cancer; Myeloid leukemia,; Lymphocyltic leukemia;
Lymphoma; Hepatic tumors; Medullary thyroid carcinoma; Multiple
myeloma; Melanoma retinoblastoma; Sarcomas of the soft tissue and
bone; Hypercalcemia including hypercalcemia associated with
malignancy; Osteolytic bone metastases and bone tumors; prevention
of bone complications related to malignant osteolysis; Osteolytic
lesions of multiple myeloma, fibrous dysplasia; pediatric
osteogenesis imperfecta; hypercalcemia, urethral (urinary tract)
malignancies, reflex sympathetic dystropy synodrome, acute back
pain after vertebral crush fracture, chronic inflammatory joint
disease, renal bone disease, extrosseous calcifications, analgesic,
vitamin D intoxication, periarticular ossifications BIBN4096BS-(1-
Anti-migraine; calcitonin gene-related Piperidinecarboxamide.
N-[2-[[5- peptide antagonist amino-1-[[4-(4-pyridinyl)-1-
piperazinyl)carbonyl]pentyl]amino]-1-
[(3,5-dibromo-4-hydroxypheny)methyl]-
2-oxoethyl]-4(1,4-dihydro-2-oxo-3(2H0-
quinazolinyl)-.[R-(R*,S*)]-); Calcitonin, including Salmon, Eel,
Osteoporosis; Diseases of the bone; bone Porcine and Human; pain;
analgesic (including pain associated with osteoporosis or cancer)
Cholecystokinin (CCK) and CCK Obesity Agonists including CCK-8;
Cromolyn Sodium (Sodium Or Disodium Asthma; Allergies
Chromoglycate); CPHPC; Reduction of amyloid deposits and systemic
amyloidoisis often (but not always) in connection with Alzheimer's
disease,Type II diabetes, and other amyloid-based diseases
Cyclosporine; Transplant Rejection Desferrioxamine (DFO); Iron
Overload Dipeptidyl peptidase IV (DPP-4) Diabetes; improving
glycemic control (e.g. inhibitors; treating hypoglycemia), obesity
Erythropoietin; Anemia Exedin and Exedin Agonists, including
Diabetes; Obesity Exendin-3 and Exendin-4; Filgrastim Reduce
infection in chemotherapy patients Follicle Stimulating Hormone
Regulate reproductive function (recombinant and natural); Gallium
nitrate; Osteoporosis; Paget's disease; Inhibits osteoclasts;
Promotes osteoblastic activity, hypercalcemia, including cancer
related hypercalcemia, urethral (urinary tract) malignancies;
anti-tumors, cancers, including urethral and bladder cancers;
lymphoma; malignancies (including bladder cancer); leukemia;
management of bone metastases (and associated pain); muliple
myeloma, attenuate immune response, including allogenic transplant
rejections; disrupt iron metabolism; promote cell migration; wound
repair; to attenuate or treat infectious processes of mycobacterium
species, including but not limited to mycobacterium tubercolosis,
and mycobacterium avium complex Glucagon; Improving glycemic
control (e.g. treating hypoglycemia and controlling hypoglycemic
reactions); obesity; a diagnostic aid in the radiogical examination
of the stomach, duodenum, small bowel and colon; treat acute
poisoning with cardiovascular agents including, but not limited to,
calcium channel blockers and beta blockers Glucagon-Like Peptide 1
(GLP-1), Diabetes; Obesity Glucagon, and Glucagon-Like Peptide 2
(GLP-2); Glucocerebrosidase; Gaucher disease (to metabolize
lipoprotein) Gonadotropin Releasing Hormone; Ovulatory dysfunction
(to stimulate ovulation) Growth Hormone Releasing Factor; Growth
Disorders Growth Hormone Releasing Hormones; Growth Disorders
Growth hormones, including human Growth Disorders growth hormones
(hGH), recombinant human growth hormones (rhGH), bovine growth
hormones, and porcine growth hormones; Heparin, including
unfractionated Thrombosis; prevention of blood coagulation heparin,
heparinoids, dermatans, chondroitins, low molecular weight heparin,
very low molecular weight heparin ultra low molecular weight
heparin and synthetic heparins including fondiparinux; Insulin,
including porcine, bovine, Diabetes; insulin resistance syndrome
human, and human recombinant, optionally having counter ions
including zinc, sodium, calcium and ammonium; Insulin-Like Growth
Factor, including Diabetes IGF-1; Interferons, including .alpha.
(e.g., interferon Viral infection, including chronic cancer and
Alfacon-1 (available as Infergen .RTM. from multiple sclerosis
Intermune, Inc. of Brisbane, Ca)), .beta., omega and .gamma.;
Interleukin-1; Interleukin-2; Interleukin- Viral Infection; Cancer
11; Interleukin-21; Leutinizing Hormone and Leutinizing Regulate
Reproductive Function Hormone Releasing Hormone; Leptin (OB
Protein); Obesity Methyphenidate salt; ADHD, Attention Deficit
Disorder, Dementia, ADDS Dementia Complex, cognitive decline in
HIV-AIDS Monoclonal Antibodies including To prevent graft
rejection; cancer Retuxin, TNF-alpha soluble receptors; Oxytocin;
Labor dysfunction (to stimulate contractions) Parathyroid Hormone
(PTH), including Osteoporosis; diseases of the bone its fragments,
including PTH 1-34 and PTH 1-38; Peptide YY (PYY) including PYY
Obesity; Diabetes; Eating Disorders; Insulin Agonists and Fragment
3-36; Resistance Syndrome Prostaglandins; Hypertension Protease
Inhibitors; Aids Somatostatin; Bleeding ulcer; erosive gastritis;
variceal bleeding; diarrhea; acromegaly; TSH- secreting pituitary
adenomas; secretory pancreatic tumors; carcinoid syndrome; reduce
proptosis/thyroid-associated ophthalmopathy; reduce macular
edema/retinopathy Thrombopoietin; Thrombocytopenia Vancomycin;
Treat or prevent antimicrobial-induced infections including, but
not limited to methacillin-resistant Staphalococcus aureus and
Staph. epidermiditis Vasopressin; Bed-wetting; antidiuretic
Vitamins; and Vitamin deficiencies Vaccines including those against
Anthrax Prevent and minimize disease or Y. Pestis, Influenza, and
Herpes.
Controlled or Sustained Release Formulations
[0172] The solid dosage forms of the present invention may be
formulated so as to prevent or retard break down in the stomach.
Controlled release formulations suitable for use in the present
invention may, for example, include an enteric coating or may be
formulated to erode from the surface.
[0173] According to one embodiment, the solid oral dosage forms
comprises a therapeutically effective amount of an active agent and
a delivery agent, wherein the solid oral dosage form has a
disintegration time of about 250 seconds to about 650 seconds when
orally administered. In another embodiment, the disintegration time
is about 350 to about 550 seconds when orally administered. In one
embodiment the disintegration time is greater than 60 seconds when
orally administered. In another embodiment, the disintegration time
is greater than 400 seconds when orally administered.
Disintegration time can be determined in water at 37.+-.2.degree.
C. using the method described in USP <701>.
[0174] The solid dosage forms of the present invention may be
covered by an enteric coating. The enteric coating may serve as the
primary control for delaying the release of the drug composition or
compositions in the solid dosage form. The enteric coating stays
intact in the stomach and prevents or retards release into the
stomach in the solid dosage form. Release of the active agent is
delayed until the solid dosage form reaches the intestine. Once in
the intestine, the higher pH causes release of the active agent.
Enteric coatings include, but are not limited to, hydroxypropyl
methylcellulose phthalate, hydroxypropyl methylcellulose acetate
succinate, polyvinyl acetate phthalate, cellulose acetate
trimellitate, cellulose acetate phthalate, poly(methacrylic
acid-ethylacrylate), and poly(methacrylic acid-methyl
methacrylate). Other enteric coatings which may be used in
accordance with the present invention are described in U.S. Pat.
No. 5,851,579, which is hereby incorporated by reference.
[0175] In one embodiment of the present invention, the enteric
coating is applied to the entire tablet, or other dosage form. In
one embodiment the enteric coating is applied to a
multi-particulate system, such as a system comprising
microparticles and/or nanoparticles discussed above.
[0176] The solid dosage forms of the present invention may be
formulated to erode from the surface of the tablet (or other dosage
uniform), or at the surface of the multi-particulate system (e.g. a
system comprising microparticles discussed above). These surface
erosion formulations slowly dissolve from the surface rather than
disintegrate. By controlling the rate of surface erosion, release
of the active agent and drug composition of the solid dosage form
can be delayed. The surface erosion formulations can be formulated
such that substantial release of the active agents or drug
compositions do not occur until the solid oral dosage form reaches
the intestines.
Enzyme Inhibiting Agents
[0177] The solid dosage forms of the present invention (comprising
the microparticles or nanoparticles of the present invention and/or
having the disintegration times discussed above) may also include
enzyme inhibiting agents. Enzyme inhibiting agents incorporated
into the solid dosage unit forms may prevent the breakdown of
insulin or other active agents that may be sensitive to enzymatic
degradation. Enzyme inhibiting agents are described in U.S. Pat.
No. 6,458,383 which is hereby incorporated by reference.
[0178] Generally, inhibitory agents can be divided into the
following classes: inhibitors that are not based on amino acids,
including P-aminobenzamidine, FK-448, camostat mesylate and sodium
glycocholate; amino acids and modified amino acids, including
aminoboronic acid derivatives and n-acetylcysteine; peptides and
modified peptides, including bacitracin, phosphinic acid dipeptide
derivatives, pepstatin, antipain, leupeptin, chymostatin,
elastatin, bestatin, hosphoramindon, puromycin, cytochalasin
potatocarboxy peptidase inhibitor, and amastatin; polypeptide
protease inhibitors, including aprotinin (bovine pancreatic trypsin
inhibitor), Bowman-Birk inhibitor and soybean trypsin inhibitor,
chicken egg white trypsin inhibitor, chicken ovoinhibitor, and
human pancreatic trypsin inhibitor; complexing agents, including
EDTA, EGTA, 1,10-phenanthroline and hydroxychinoline; and
mucoadhesive polymers and polymer-inhibitor conjugates, including
polyacrylate derivatives, chitosan, cellulosics, chitosan-EDTA,
chitosan-EDTA-antipain, polyacrylic acid-bacitracin, carboxymethyl
cellulose-pepstatin, polyacrylic acid-Bowman-Birk inhibitor.
[0179] The choice and levels of the enzyme inhibitor are based on
toxicity, specificity of the proteases and the potency of
inhibition, and will be apparent to those skilled in the art.
[0180] Without wishing to be bound by theory, it is believed that
an inhibitor can function solely or in combination as: a
competitive inhibitor, by binding at the substrate binding site of
the enzyme, thereby preventing the access to the substrate
(examples of inhibitors believed to operate by this mechanism are
antipain, elastatinal and the Bowman Birk inhibitor); a
non-competitive inhibitor that can be simultaneously bound to the
enzyme site along with the substrate, as their binding sites are
not identical; and/or a complexing agent due to loss in enzymatic
activity caused by deprivation of essential metal ions out of the
enzyme structure.
Solid Pharmaceutical Composition Embodiment
[0181] This application hereby incorporates by reference
International Publication No. WO 2005/004900 and its priority
document U.S. Provisional Application No. 60/486,495, filed Jul.
11, 2003, in their entireties.
[0182] The pharmacologically active agents suitable for use in the
solid pharmaceutical composition of the instant invention include
both therapeutic as well as preventative agents and is directed
particularly to agents which by themselves do not pass or which
pass only a small amount of the administered dose through the
gastro-intestinal mucosa and/or are susceptible to cleavage by
acids and enzymes in the gastro-intestinal tract. The
pharmacologically active agents include, but are not limited to
proteins; polypeptides; hormones; polysaccharides including
mixtures of muco-polysaccharides; carbohydrates; lipids; and
combinations thereof.
[0183] Specific examples of pharmacologically active agents
include, but are not limited to, the following, including
synthetic, natural or recombinant sources thereof: growth hormone,
including human growth hormones (hGH), recombinant human growth
hormones (rhGH), bovine growth hormones, and porcine growth
hormones; growth hormone-releasing hormones; interferons, including
.alpha., .beta., and .gamma.-interferon; interleukin-1;
interleukin-2; insulin, including porcine, bovine, human, and human
recombinant, optionally having counter ions including sodium, zinc,
calcium and ammonium; insulin-like growth factor, including IGF-1;
heparin, including unfractionated heparin, heparinoids, dermatans,
chondroitins, low, very low and ultra low molecular weight heparin;
calcitonin, including salmon, porcine, eel, chicken and human;
erythopoietein; atrial naturetic factor; antigens; monoclonal
antibodies; somatostatin; protease inhibitors; adrenocorticotropin,
gonadotropin releasing hormone; oxytocin;
leutinizing-hormone-releasing hormone; follicle stimulating
hormone; glucocerebrosidase; thrombopoietin; filgrastim;
prostaglandins; cyclosporin; vasopressin; cromolyn sodium (sodium
or disodium chromoglycate); vancomycin; desferrioxamine (DFO);
parathyroid hormone (PTH), including its fragments; antimicrobials,
including anti-fungal agents; vitamins; analogs, fragments,
mimetics or polyethylene glycol (PEG)-modified derivatives of these
compounds; or any combination thereof.
[0184] An interesting pharmacologically active agent is a
pharmacologically active peptide, particularly bone active agents,
and even more particularly calcitonin.
[0185] Bone active agents include classes of agents which display
in vivo pharmacological activity in animals such as stabilization,
healing, or growth of bone, deceleration or inhibition of bone
turnover, deceleration or inhibition of bone resorption, inhibition
of osteoclast activity, and stimulation of osteoblast activity.
Some of these agents may be peptidic, for example calcitonins,
parathyroid hormone (PTH), PTH fragments, analogs and releasers,
and Transforming Growth Factors (TGFs) fragments, analogs and
releasers. The bone active agents may also be small molecule
non-peptidic structures which show in vivo pharmacological bone
activities as described above in this paragraph.
[0186] A known class of such pharmacologically active agents,
calcitonins, have varying pharmaceutical utility and are commonly
employed in the treatment of e.g. Paget's disease, hypercalcemia
and postmenopausal osteoporosis. Various calcitonins, including
salmon, pig and eel calcitonin are commercially available and
commonly employed for the treatment of e.g. Paget's disease,
hypercalcemia of malignancy and osteoporosis. The calcitonin can be
any calcitonin, including natural, synthetic or recombinant sources
thereof, as well as calcitonin derivatives such as 1, 7-Asu-eel
calcitonin. The compositions can comprise a single calcitonin or
any combination of two or more calcitonins. The preferred
calcitonin is synthetic salmon calcitonin.
[0187] The calcitonins are commercially available or may be
synthesized by known methods.
[0188] The amount of pharmacologically active agent is generally an
amount effective to accomplish the intended purpose, e.g. a
therapeutically effective amount. However, the amount can be less
than that amount when a plurality of the compositions are to be
administered, i.e., the total effective amount can be administered
in cumulative dosage units. The amount of active agent can also be
more than the effective amount when the composition provides
sustained release of the pharmacologically active agent. The total
amount of active agent to be used can be determined by methods
known to those skilled in the art. However, because the
compositions may deliver the active agent more efficiently than
prior compositions, less amounts of active agent than those used in
prior dosage unit forms or delivery systems can be administered to
a subject while still achieving the same blood levels and/or
therapeutic effects.
[0189] When the pharmacologically active agent is salmon
calcitonin, the appropriate dosage will, of course, vary depending
upon, for example, the host and the nature and severity of the
condition being treated. However, in general, satisfactory results
will be obtained systemically at daily dosages of from about 0.5
g/kg to about 10 g/kg animal body weight, preferably 1 g/kg to
about 6 body weight.
[0190] The pharmacologically active agent generally comprises from
0.05 to 70 percent by weight relative to the total weight of the
overall pharmaceutical composition, preferably an amount of from
0.01 to 50 percent by weight, more preferably 0.3 to 30 percent by
weight relative to the total weight of the overall pharmaceutical
composition.
[0191] The pharmaceutically acceptable inactive excipients may
include polymers and inactive compounds which for example, aid the
formulation or manufacturing of the solid oral dosage form
contemplated by the present invention or which may aid the release
of the solid oral composition in the gastro-intestinal
environment.
[0192] The pharmaceutical inactive ingredients, referred to above,
for example optionally include crospovidones and povidones, which
may be any crospovidone and povidone. Crospovidone is a synthetic
crosslinked homopolymer of N-vinyl-2-pyrrolidone, also called
1-ethenyl-2-pyrrolidinone, having a molecular weight of 1,000,000
or more. Commercially available crospovidones include Polyplasdone
XL, PolyplasdoneXL-10, Polyplasdone INF-10 available from ISP,
Kollidon CL, available from BASF Corporation. The preferred
crospovidone is Polyplasdone XL.
[0193] Povidone is a synthetic polymer consisting of linear
1-vinyl-2-pyrrolidinone groups having a molecular weight generally
between 2,500 and 3,000,000. Commercially available povidones
include Kollidon K-30, Kollidon K-90F available from BASF
Corporation and Plasdone K-30 and Plasdone K-29/32, available from
ISP.
[0194] As mentioned above, the crospovidones and povidones are
commercially available. Alternatively, they may be synthesized by
known processes.
[0195] The crospovidone, povidone or combination thereof is
generally present in the compositions in an amount of from 0.5 to
50 percent by weight relative to the total weight of the overall
pharmaceutical composition, preferably an amount of from 2 to 25
percent, more preferably 5 to 20 percent by weight relative to the
total weight of the pharmaceutical composition.
[0196] The delivery agents useful in the solid pharmaceutical
composition are any agents useful for delivering the particular
pharmacologically active agent. Suitable delivery agents are any
one of the 123 modified amino acids disclosed in U.S. Pat. No.
5,866,536 or any one of the 193 modified amino acids described in
the U.S. Pat. No. 5,773,647 or any combination thereof. The
contents of the aforementioned U.S. Pat. Nos. 5,773,647 and
5,866,536 are hereby incorporated by reference in their entirety.
In addition, the delivery agent can be the disodium salt of any of
the aforementioned modified amino acids as well as ethanol solvates
and hydrates thereof. Suitable compounds include compounds of the
following formula I
##STR00006##
wherein:
[0197] R.sup.1, R.sup.2, R.sup.3, and R.sup.4 are independently
hydrogen, --OH, --NR.sup.6R.sup.7, halogen, C.sub.1-C.sub.4 alkyl,
or C.sub.1-C.sub.4 alkoxy;
[0198] R.sup.5 is a substituted or unsubstituted C.sub.2-C.sub.6
alkylene, substituted or unsubstituted C.sub.2-C.sub.16 alkenylene,
substituted or unsubstituted C.sub.1-C.sub.12 alkyl(arylene), or
substituted or unsubstituted aryl(C.sub.1-C.sub.12 alkylene);
and
[0199] R.sup.6 and R.sup.7 are independently hydrogen, oxygen, or
C.sub.1-C.sub.4 alkyl; and hydrates and alcohol solvates thereof.
The compounds of formula I as well as their disodium salts and
alcohol solvates and hydrates thereof are described in WO
00/059863, along with methods for preparing them.
[0200] The disodium salt may be prepared from the ethanol solvate
by evaporating or drying the ethanol solvate by methods known in
the art to form the anhydrous disodium salt. Drying is generally
carried out at a temperature of from about 80 to about 120.degree.
C., preferably from about 85 to about 90.degree. C., and most
preferably at about 85.degree. C. The drying step is generally
performed at a pressure of 26'' Hg or greater. The anhydrous
disodium salt generally contains less than about 5% by weight of
ethanol and preferably less than about 2% by weight of ethanol,
based on 100% total weight of anhydrous disodium salt.
[0201] The disodium salt of the delivery agent can also be prepared
by making a slurry of the delivery agent in water and adding two
molar equivalents of aqueous sodium hydroxide, sodium alkoxide or
the like. Suitable sodium alkoxide include, but are not limited to,
sodium methoxide, sodium ethoxide, and combinations thereof.
[0202] A still further method of preparing the disodium salt is by
reacting the delivery agent with one molar equivalent of sodium
hydroxide to yield the disodium salt.
[0203] The disodium salt can be isolated as a solid by
concentrating the solution containing the disodium salt to a thick
paste by vacuum distillation. This paste may be dried in a vacuum
oven to obtain the disodium salt of the delivery agent as a solid.
The solid can also be isolated by spray drying an aqueous solution
of the disodium salt.
[0204] The delivery agents may be prepared by methods known in the
art, e.g., as mentioned above, by methods described in U.S. Pat.
Nos. 5,773,647 and 5,866,536.
[0205] The ethanol solvates, as described in the aforementioned
International Publication No. WO 00/059863, include, but are not
limited to, a molecular or ionic complex of molecules or ions of
ethanol solvent with molecules or ions of the disodium salt of the
delivery agent. Typically, the ethanol solvate contains about one
ethanol molecule or ion for every molecule of disodium salt of the
delivery agent.
[0206] The ethanol solvate of the disodium salt of the delivery
agent can be prepared by dissolving the delivery agent in ethanol.
Typically, each gram of delivery agent is dissolved in from about 1
to about 50 mL of ethanol and generally, from about 2 to about 10
mL of ethanol. The delivery agent/ethanol solution is then reacted
with a molar excess of a sodium containing salt, such as a
monosodium containing salt, relative to delivery agent, i.e. for
every mole of delivery agent there is more than one mole of sodium
cations, yielding the ethanol solvate. Suitable monosodium salts
include, but are not limited to, sodium hydroxide; sodium
alkoxides, such as sodium methoxide and sodium ethoxide; and any
combination of the foregoing. Preferably, at least about two molar
equivalents of the monosodium containing salt are added to the
ethanol solution, i.e. for every mole of delivery agent there is at
least about two moles of sodium cations. Generally, the reaction is
performed at or below the reflux temperature of the mixture, such
as at ambient temperature. The ethanol solvate is then recovered by
methods known is the art, such as, concentration of the resulting
slurry at atmospheric distillation, cooling the concentrated slurry
and filtering the solid. The recovered solid can then be vacuum
dried to obtain the ethanol solvate.
[0207] The hydrates of the disodium salts of the delivery agents
may be prepared by drying the ethanol solvate to form an anhydrous
disodium salt, as described above, and hydrating the anhydrous
disodium salt. Preferably, the monohydrate of the disodium salt is
formed. Since the anhydrous disodium salt is very hydroscopic, the
hydrate forms upon exposure to atmospheric moisture. Generally, the
hydrating step is performed at from about ambient temperature to
about 50.degree. C., preferably ambient temperature to about
30.degree. C. and in an environment having at least 50% relative
humidity. Alternatively, the anhydrous disodium salt may be
hydrated with steam.
[0208] The preferred delivery agents for the solid pharmaceutical
composition are N-(5-chlorosalicyloyl)-8-aminocaprylic acid (5-CNAC
acid), N-(10-[2-hydroxybenzoyl]-amino) decanoic acid (SNAD acid),
N-(8-[2-hydroxybenzoyl] amino) caprylic acid (SNAC acid) and their
monosodium and disodium salts, ethanol solvates of their sodium
salts and the monohydrates of their sodium salts and any
combinations thereof. The most preferred delivery agent is the
disodium salt of 5-CNAC acid and the monohydrate thereof.
[0209] The delivery agents 5 CNAC acid, SNAD acid, and SNAC acid
(and their salts) are very water soluble and nearly fully, i.e.
greater than 90%, absorbed by the gastro-intestinal tract whether
it is ingested in micronized or coarse form. However, when a
micronized form of one of these carrier agents is employed in the
composition, the absorption of the pharmacologically active agent
of the present composition is more completely absorbed into the
blood stream.
[0210] A micronized form of the delivery agent, which is utilized
in preparation of the solid pharmaceutical composition or solid
oral dosage form of the present invention, is defined as a delivery
agent which, when added to the present composition mixture of
pharmacologically active agent and pharmaceutical inactive
ingredients, has an average particle size of less than 40
micrometers. Desirably the delivery agent of the present invention
has a micronized form which is defined as an average particle size
of less than 20 microns. More interestingly, the delivery agent for
the present invention has a micronized form which is defined as an
average particle size of less than 10 microns.
[0211] Micronized forms of the delivery agent of the present
invention may be prepared by grinding it in a grinding mill which
is acceptable for grinding pharmaceutical ingredients and which is
capable of grinding the pharmaceutical ingredients and/or delivery
agent to a fine and uniform micronized particle size. An example of
such a grinding mill is an Air Jet Mill GemT & commat; (Copley
Scientific, Ltd., Nottingham, UK). The finely ground delivery agent
either separately or finely ground delivery agent plus any
combination of finely ground additional ingredients of the present
invention may then be screened, for example, over a mesh screen
having the appropriate openings, in order to allow only those
ingredients which have the required particle size to pass through
and be collected for use in the present invention.
[0212] The solid pharmaceutical compositions typically contain a
delivery effective amount of one or more of the delivery agents,
i.e. an amount sufficient to deliver the active agent for the
desired effect. Generally, the delivery agent is present in an
amount of 2.5% to 99.4% by weight, more preferably 25% to 50% by
weight.
[0213] The solid pharmaceutical compositions may be provided as a
capsule including a soft-gel capsule, tablet, caplet or other solid
oral dosage form, all of which can be prepared by methods well
known in the art.
[0214] The solid pharmaceutical compositions may additionally
comprise additives in amounts customarily employed including, but
not limited to, a pH adjuster, a preservative, a flavorant, a
taste-masking agent, a fragrance, a humectant, a tonicifier, a
colorant, a surfactant, a plasticizer, a lubricant such as
magnesium stearate, a flow aid, a compression aid, a solubilizer,
an excipient, a diluent such as microcrystalline cellulose, e.g.
Avicel PH 102.RTM. (supplied by FMC Corporation, 1735 Market Street
Philadelphia, Pa. 19103, USA), or any combination thereof. Other
additives may include phosphate buffer salts, citric acid, glycols,
and other dispersing agents.
[0215] The solid pharmaceutical composition may also include one or
more enzyme inhibitors, such as actinonin or epiactinonin and
derivatives thereof; aprotinin, Trasylol andBowman-Birk
inhibitor.
[0216] Further, a transport inhibitor, i.e. a p-glycoprotein such
as Ketoprofin, may be present in the compositions of the present
invention.
[0217] Preferably, the solid pharmaceutical compositions include a
diluent, such as Avicel.RTM., and a lubricant, such as magnesium
stearate.
[0218] The solid pharmaceutical compositions can be prepared by
first grinding either the delivery agent or the delivery agent with
any combination of the additional ingredients of the present
composition to a micronized particle size. The micronized delivery
agent or micronized delivery agent plus micronized additional
ingredients of the present invention may then be further processed
by conventional methods e.g. by blending a mixture of the active
agent or active agents, the delivery agent, the crospovidone or
povidone and other ingredients, kneading, and filling into capsules
or, instead of filling into capsules, molding followed by further
tableting or compression-molding to give tablets. In addition, a
solid dispersion may be formed by known methods followed by further
processing to form a tablet or capsule.
[0219] Preferably, the ingredients in the solid pharmaceutical
compositions are homogeneously or uniformly mixed throughout the
solid dosage form.
[0220] The solid pharmaceutical compositions may be administered to
deliver an active agent to any animal in need thereof, including,
but not limited to, mammals, such as rodents, cows, pigs, dogs,
cats, and primates, particularly humans.
EXAMPLES
[0221] The following examples illustrate the invention without
limitation. All parts are given by weight unless otherwise
indicated.
Example 1
1. Test Articles
[0222] a. Co-Processed Insulin/Delivery Agent Microparticles Used
for Site Specific, In Situ Experiment and Oral Gavage
Experiments
[0223] Recombinant human zinc insulin (50 mg) and sodium 4-CNAB
(7.5 g) were dissolved in 50 ml of deionized water. The clear
solution was dried with nitrogen flow at room temperature for 24
hours. The obtained coprocessed cake was milled into fine
particles, which were then sieved through a 40/60 mesh screen to
obtain microparticles of a specific size range. The size of the
microparticles used in the current study ranged from 250 to 420 m.
These microparticles contained by weight 0.55% of insulin, 9.5% of
water and 89.5% of delivery agent. A total of approximately 90%
(w/w) of insulin was recovered from this process.
[0224] Particles were measured by passing them through seives with
different size openings (850 .mu.m, 425 .mu.m, 250 .mu.m, 150
.mu.m, 45 .mu.m). With this method, it can be determined that the
median particle size ranges from about 45 to about 850 .mu.m, from
about 45 to about 150 .mu.m, from about 150 to about 250 .mu.m,
from about 250 to about 425 .mu.m, or from about 425 to about 850
.mu.m.
[0225] Insulin content in the microparticles was measured with
reversed phase HPLC (Phenomenex column: Luna 3u C18 (2) 100 .ANG.,
150.times.4.6 mm, 3 micro; mobile phases: A, 0.1% TFA in water; B,
0.1% TFA in acetonitrile; Detector: UV280 nm). Water contents of
the particles were measured with a 737 KF coulometer.
[0226] b. Capsules Loaded with the Microparticles for Oral
Gavage
[0227] Gelatin capsules (size #9) were used in the rat studies. The
necessary amount of microparticles loaded manually into the gelatin
capsules were determined based on an average rat body weight of 350
mg. Each loaded capsule contained approximately 16 mg of the
microparticles (equivalent to 0.0875 mg of insulin).
[0228] c. Insulin/Delivery Agent Mini-Tablets for Oral Gavage
Experiment
[0229] Insulin was well mixed with delivery agent at a ratio of
1:150 (w/w), which corresponded to 0.67% (w/w) of insulin. Based on
an average rat body weight of 350 mg, a total amount of 26.43 mg of
the mixed powder, which contained 0.175 mg of insulin and 26.26 mg
of delivery agent, was directly compressed into tablets under a
pressure of 1000 psi in a Carver press. The cylindrical
mini-tablets were 2 mm in diameter and 6 mm in height.
[0230] d. Capsules Loaded with Insulin/Delivery Agent Physical
Blend for Oral Gavage Experiment
[0231] Insulin was well mixed with delivery agent at a ratio of
1:150 (w/w). The amount of insulin and delivery agent mixture
loaded manually into the gelatin capsules (size #9) were determined
based on an average rat body weight of 350 mg. Each capsule
contained 26.43 mg of the mixture (equivalent to 0.175 mg
insulin).
2. Direct Dosing Procedures for In Situ Experiments
[0232] A schematic of the direct dosing procedure is shown in FIG.
1. Surgery was carried out in a clean environment using a clean lab
coat, mask, safety goggle, gloves and surgical cap. Anesthesia was
induced to the Sprague Dawley rats with 5% isoflurane as an
induction concentration, and maintained at 2% isoflurane in pure
oxygen to the completion of the study.
[0233] a. Stomach Direct Dosing
[0234] After the right jugular vein was catheterized for sampling
blood, the skin over the esophagus and trachea was dissected, and
the musculus digastricul venter rostralis (protective muscular
bundles) was identified and dissected to make an access toward the
esophagus. The esophagus was partially severed, and inserted with a
12 cm PE204 tubing for a segment of the esophagus measuring 6-9 cm.
The dosing formulation was introduced through this tubing using a
blunt wire to push in the microparticles. After dosing, the
esophagus was ligated with a 3-0 silk suture for preventing any
leakage from the stomach.
[0235] b. Jejunum Direct Dosing
[0236] After the right jugular catheterization for the blood
sampling, the abdominal cavity was opened by dissecting the linea
alba toward the sternum, thus exposing the xiphoid cartilage. The
most proximal segment ofjejunum was first identified. A less
vascularized section of the proximal jejunum was partially nipped,
and a dosing tube was introduced toward the distal end. After
dosing, the dosing tube was removed, and a 2 cm PE206 tubing was
pushed in, and placed so that the nipped wound was located in the
middle of both ends of the 2 cm tubing. A suture was tied around
the tubing with jejunum at both ends, and the wound was closed with
a drop of a Vetbond.TM. tissue adhesive (available from 3M of St.
Paul, Minn.).
3. Oral Gavage-Procedures
[0237] Studies were carried out in Sprague Dawley rats (body weight
was approximately 350 grams) by oral gavage administration. The
mini tablets or capsules were administrated orally in rats using a
modified gavage tubing with a trocar. Rats were fasted for about 24
hours and anesthetized by intramuscular administration of ketamine
(44 mg/kg) and thorazine (1.5 mg/kg). At pre-determined time
intervals, blood samples were drawn from the tail artery and were
appropriately prepared as either plasma or serum for glucose and
insulin bioassays. The animal was sacrificed at the end of the
experiment and rat gastrointestinal mucosa was observed for any
sign of local toxicity.
4. Bioassay Procedures
[0238] Rat serum concentrations of insulin were determined using
Insulin ELISA Test Kit (DSL Inc.). The limit of quantitation (LOQ)
has been established at 12.5 .mu.U/mL, with the calibrated linear
range of the assay up to 250 .mu.U/mL. Changes in blood glucose
levels were measured using a glucometer.
5. Results
[0239] a. Site Specific Study (In Situ) Results
[0240] The concentration of insulin and the change in glucose level
following direct dosing of the coprocessed microparticles to the
stomach and the jejunum are shown in FIGS. 2 and 3, respectively.
The individual data are listed in Tables 2 to 5.
[0241] Insulin concentration from dosing to the jejunum reached a
maximum value at the first sampling point (t.sub.max.ltoreq.15 min)
from each formulation. The corresponding t.sub.min of glucose
occurred approximately 30 min. later.
TABLE-US-00002 TABLE 2 Direct dosing of coprocessed microparticles
to the stomach 1) Insulin Insulin Stomach Time Rat# (min) 1 #2 #3
#4 #5 #6 #7 #8 mean SD SEM CV 0 12.5 12.5 12.5 12.5 12.5 12.5 12.5
12.9 0.0 0.0 0.0 15 114.5 72.8 80.9 12.6 210.0 12.5 118.7 158.5
97.6 68.1 24.1 69.8% 30 95.3 19.3 35.5 12.5 211.0 12.5 15.1 66.0
58.3 68.4 24.2 117.2% 45 62.8 12.5 894.0 12.5 213.0 12.5 15.0 12.5
154.3 306.7 108.5 198.8% 60 18.2 12.5 157.0 12.5 174.0 140.3 12.5
12.5 67.4 74.7 26.4 110.9% 90 12.5 12.5 12.5 12.5 61.3 12.5 12.5
74.2 26.3 25.8 9.1 98.1% AUC.sub.0.fwdarw.90 4780 2132 18970 1127
14438 4001 2795 5046 6661 6451 2281 96.8% 2) Glucose Change from
base line Stomach Time Rat# (min) 1 #2 #3 #4 #5 #6 #7 #8 Mean SD
SEM CV 0 0 0 0 0 0 0 0 0 0 0 0 15 -9.9 -21.0 -12.6 -38.1 -6.6 -24.8
3.0 -13.9 -15.5 12.5 4.4 -80.6% 30 -44.3 -43.8 -25.8 -56.1 -9.8
-54.4 -3.4 -47.6 -35.7 20.2 7.1 -56.6% 45 -75.0 -50.9 -30.2 -73.6
-17.1 -73.6 -14.8 -64.2 -49.9 25.8 9.1 -51.6% 60 -80.7 -51.3 -31.1
-66.1 -18.7 -72.8 -21.9 -55.1 -49.7 23.5 8.3 -47.3% 90 -62.3 -32.1
-35.2 -59.7 -20.3 -63.6 -29.5 -28.3 -41.4 17.5 6.1 -42.3% (two rat
data were removed, rat #1-2 stomach) *Insulin (0.5 mg/kg), Delivery
Agent (75 mg/kg))
TABLE-US-00003 TABLE 3 Direct dosing of coprocessed microparticles
to the jejunum data 1) Insulin Insulin Jejunum Time (min) #9 #10
#11 #12 #13 #14 #15 #16 mean SD SEM CV 0 12.5 12.5 12.5 12.5 12.5
12.5 12.5 12.5 0.0 0.0 0.0 15 413.2 1193.3 669.4 1177.5 2270.6
228.9 954.4 374.9 910.3 661.1 233.8 72.6% 30 354.3 148.4 70.5 64.7
481.0 168.4 782.9 57.4 265.9 258.2 91.3 97.1% 45 79.5 28.0 20.5
26.5 170.8 148.6 531.6 12.5 127.3 174.3 61.6 137.0% 60 23.1 14.7
12.5 16.8 71.6 117.0 200.1 12.5 58.5 68.4 24.2 116.9% 90 12.5 12.5
12.5 12.5 30.8 12.5 37.5 12.5 17.9 10.2 3.6 56.8%
AUC.sub.0.fwdarw.90 13506 21158 11969 19690 46003 11102 39192 7235
21232 14054 4969 66.2% 2) Glucose Change from base line Jejunum
Time (min) #9 #10 #11 #12 #13 #14 #15 #16 Mean SD SEM CV 0 0 0 0 0
0 0 0 0 0 0 0 15 -50.8 -35.6 -16.7 -16.0 -61.1 -38.2 -62.8 -36.3
-39.7 17.9 6.3 -45.0% 30 -67.3 -65.5 -59.5 -35.8 -81.8 -62.5 -78.1
-52.9 -62.9 14.4 5.1 -22.9% 45 -64.4 -68.5 -76.3 -56.4 -74.4 -71.6
-78.1 -53.3 -67.9 9.2 3.2 -13.5% 60 -62.7 -62.5 -71.3 -62.7 -62.1
-65.6 -74.0 -42.2 -62.9 9.5 3.4 -15.1% 90 -62.4 -49.8 -69.7 -55.6
-41.8 -44.2 -69.4 -26.7 -52.5 14.9 5.3 -28.3% (two rat data were
removed, rat# 2 jejunum and rat# 4 jejunum) (Insulin (0.5 mg/kg),
Delivery Agent (75 mg/kg))
TABLE-US-00004 TABLE 4 Direct dosing of coprocessed microparticles
to the stomach 1) Insulin stomach Time (min) sto-1 sto-2 sto-3
sto-4 sto-5 sto-6 sto-7 sto-8 mean SD SEM CV 0 12.5 12.5 41.2 21.5
12.5 12.5 12.5 12.5 17.2 9.5 3.4 55.4% 15 12.5 20.8 14.7 12.5 12.5
12.5 26.4 12.5 16.0 4.9 1.7 30.7% 30 12.5 12.5 12.5 12.5 12.5 12.5
12.5 12.5 12.5 0.0 0.0 0.0% 45 48.8 12.5 12.5 12.5 12.5 12.5 12.5
12.5 17.0 12.0 4.3 70.4% 60 12.5 12.5 12.5 12.5 12.5 12.5 12.5 12.5
12.5 0.0 0.0 0.0% 90 12.5 12.5 12.5 12.5 12.5 12.5 12.5 12.5 12.5
0.0 0.0 0.0% AUC.sub.0.fwdarw.90 1670 1250 1373 1193 1125 1125 1334
1125 1274 187 66.0 14.6& 2) Glucose Stomach Change from base
line Rat# Time (min) 1 #2 #3 #4 #5 #6 #7 #8 mean SD SEM CV 0 0 0 0
0 0 0 0 0 0 0 0.0 15 -2.2 -12.3 2.2 0.8 -2.2 -12.3 2.2 0.8 -2.9 5.7
2.0 -196.5% 30 -14.4 -10.1 -6.5 0.8 -14.4 -10.1 -6.5 0.8 -7.6 5.6
1.9 -73.7% 45 -15.8 -8.8 -12.7 0.0 -15.8 -8.8 -12.7 0.0 -9.3 5.9
2.1 -63.4% 60 -15.8 -11.4 -17.9 -5.8 -15.8 -11.4 -17.9 -5.8 -12.7
4.6 1.6 -36.2% 90 -19.1 -16.3 -8.6 -6.6 -19.1 -16.3 -8.6 -6.6 -12.7
5.2 1.8 -40.9% (Insulin (0.25 mg/kg), Delivery Agent (37.5
mg/kg))
TABLE-US-00005 TABLE 5 Direct dosing of coprocessed microparticles
to the jejunum data 1) Insulin jejunum Time (min) jej-1 jej-2 jej-3
jej-4 jej-5 jej-6 jej-7 jej-8 mean SD SEM CV 0 12.5 12.5 12.5 12.5
12.5 12.5 12.5 12.5 12.5 0.0 0.0 0.0% 15 428.5 532.4 232.6 12.5
62.0 186.6 79.6 160.5 219.2 170.7 60.4 77.8% 30 67.9 100.8 44.7
12.5 15.5 12.5 14.2 49.3 39.7 30.4 10.7 76.4% 45 16.8 40.8 26.6
12.5 12.5 12.5 12.5 12.5 18.4 9.7 3.4 52.6% 60 12.5 24.7 17.3 12.5
12.5 12.5 12.5 12.5 14.6 4.1 1.5 28.1% 90 12.5 12.5 12.5 12.5 12.5
12.5 12.5 12.5 12.5 0.0 0.0 0.0% AUC.sub.0.fwdarw.90 8261 10947
5229 1125 1913 3737 2157 3897 4658 3392 1199 72.8% 2) Glucose
Change from base line Jejunum Time (min) #9 #10 #11 #12 #13 #14 #15
#16 Mean SD SEM CV 0 0 0 0 0 0 0 0 0 0 0 0.0 15 -29.7 -20.5 -35.8
1.3 -28.9 -18.2 -25.9 -25.6 -22.9 11.2 3.9 -48.9% 30 -52.2 -54.5
-41.9 -1.3 -50.0 -60.3 -36.8 -52.8 -43.7 18.7 6.5 -42.8% 45 -63.6
-65.3 -43.8 -0.7 -56.8 -69.2 -40.8 -59.7 -59.0 22.3 7.8 -37.8% 60
-56.9 -69.4 -33.8 11.6 -56.8 -62.1 -27.2 -55.7 -41.9 28.1 10.6
-67.1% 90 -63.9 -59.7 -26.9 8.3 -50.0 -31.8 -11.8 -28.4 -28.6 22.7
8.5 -79.4% (Insulin (0.25 mg/kg), Delivery Agent (37.5 mg/kg))
[0242] b. Results from Oral Gavage Experiments Using Tablet and
Capsules
[0243] The glucose and insulin data from the three formulations
tested are shown in FIGS. 4 and 5, respectively. The individual
data are listed in Tables 6 to 7. The results from the direct
dosing studies to the stomach and jejunum are included for
comparison. The individual glucose and insulin data for the simple
mix of insulin and delivery agent is shown in Table 8.
[0244] In the group of 10 rats that was dosed with capsules
containing microparticles of coprocessed insulin and carrier, the
average minimum glucose lowering was 70% from baseline at 30
minutes. One rat died at 15-30 minutes, likely due to hypoglycemia,
six rats were rescued at 30 minutes with dextrose, an additional
rat was rescued at 60 minutes, and two of the six that were rescued
at 30 minutes died after 60 minutes. There were no signs of GI
irritation or GI damage from the oral gavage procedure from
necropsies of the rats after the experiment. The average minimum
glucose lowering from tablets that contained the same amounts of
insulin and carrier was 50%.
[0245] The corresponding insulin concentrations are shown in FIG.
5. Insulin concentration is highest from the coprocessed
microparticles in a capsule, followed by the tablet and the capsule
of the simple mix.
[0246] In the oral gavage studies using capsules containing
coprocessed microparticles, two (of 10) rats were found to exhibit
high insulin absorption. Retainer samples were reassayed and
insulin levels were approximately the same as those from the
original samples, as shown in Table 6(3), shown above. Insulin
levels with and without two high responders are shown in FIGS. 6
and 7, respectively. The individual and average insulin and glucose
profiles from N=10 and N=8 are shown in FIGS. 13 to 16.
TABLE-US-00006 TABLE 6 Oral gavage of tablets: Insulin (0.5 mg/kg),
Delivery Agent (75 mg/kg) 1) Insulin Time Rat# (min) 11 #12 #13 #14
#15 #16 #17 #18 #19 #20 mean SD SEM CV 0 12.5 12.5 12.5 12.5 12.5
12.5 12.5 12.5 12.8 12.5 12.5 0.1 0.0 0.8% 15 468.8 162.1 700.1
12.5 1363.4 197.4 565.4 57.0 114.4 12.5 365.4 426.3 134.8 116.7% 30
90.5 14.5 108.6 12.5 174.0 14.4 62.7 117.5 20.0 16.1 63.1 57.2 18.1
90.7% 45 15.2 32.0 22.9 12.5 44.5 12.5 16.3 43.3 12.5 12.5 22.4
12.9 4.1 57.6% 60 12.5 12.5 13.9 12.5 23.2 16.7 12.5 12.5 12.5 12.5
14.1 3.5 1.1 24.6% 90 12.5 12.5 12.5 12.5 12.5 12.5 12.5 12.5 12.5
12.5 12.5 0.0 0.0 0.0% AUC.sub.0.fwdarw.90 9180 3692 13068 1125
24532 4022 10229 3830 2768 1179 7363 7259 2295 98.6% 2) Glucose
Change from baseline Time Rat# (min) 1 #2 #3 #4 #5 #6 #7 #8 #9 #10
mean SD SEM CV 0 0 0 0 0 0 0 0 0 0 0 0 0 0.0 0.00% 15 -51.4 -38.9
-36.5 -37.8 -23.8 -26.7 12.1 16.5 25.0 54.3 -10.7 35.0 11.1 -326.7%
30 -70.8 -69.4 -66.2 -68.3 -67.9 -68.9 -57.1 -41.8 -20.7 47.8 -48.3
37.4 11.8 -77.4% 45 -66.7 -63.9 -64.9 -57.3 -57.1 -52.2 -44.0 -27.5
-27.2 52.2 -40.9 35.7 11.3 -87.4% 60 -54.2 -47.2 -41.9 -42.7 -34.5
-33.3 -9.9 -4.4 -1.1 51.1 -21.8 31.7 10.0 -145.1% 90 -50.0 -26.4
-17.6 -19.5 -13.1 5.6 9.9 11.0 32.6 106.5 3.9 42.9 13.6 1099.9%
TABLE-US-00007 TABLE 7 Oral gavage of capsules containing
coprocessed insulin and delivery agent data 1) Insulin Time Rat#
(min) 11 #12 #13 #14 #15 #16 #17 #18 #19 #20 mean SD SEM CV 0 12.5
12.5 12.5 12.5 12.5 12.5 12.5 12.5 12.5 12.5 0.0 0.0 0.0 0.0% 15
461.1 35.2 400.0 1080.6 143.1 36.7 7970.5 1611.3 369.8 922.5 1303.1
2396.7 757.9 183.9% 30 244.0 120.0 3268.4 35.3 32.7 5915.1 201.0
150.9 240.3 1134.2 2070.4 654.7 182.5% 45 26.9 13.7 3132.7 18.9
201.2 3309.5 58.7 12.5 38.3 756.9 1399.0 442.4 184.8% 60 13.8 12.5
2129.7 12.5 28.7 3693.9 16.0 12.5 12.5 659.1 1335.7 422.4 202.6% 90
12.5 12.5 984.9 12.5 12.5 12.5 12.5 151.4 367.5 116.2 242.7%
AUC.sub.0.fwdarw.90 11752 3096 175011 3522 4986 285725 28279 8561
18579 59946 100827 33609 168.2% (n = 9) AUC.sub.0.fwdarw.90 11752
3096 3522 4986 28279 8561 18579 11254 9279 3507 82.4% (n = 7) 2)
Glucose Time (min) #11 #12 #13 #14 #15 #16 #17 #18 #19 #20 mean SD
SEM CV 0 0 0 0 0 0 0 0 0 0 0 0 0 0.0 0.0% 15 -27.9 41.6 -32.5 -30.4
-23.3 19.8 -53.9 -5.5 -19.6 -25.3 -15.7 27.7 9.8 -176.3% 30 -79.0
-17.5 -76.6 -64.0 -40.1 -78.0 -77.0 -63.8 -77.0 -63.7 21.4 7.5
-33.5% 45 -42.1 -46.4 -62.6 -36.7 -66.7 -53.9 -4.9 10.9 -50.0 -39.2
25.9 9.1 -66.2% 60 3.2 -34.9 -61.4 -13.3 -75.3 -70.3 -7.2 64.5
-48.3 -30.8 45.9 16.2 -149.0% 90 68.4 -7.2 -62.6 20.7 92.0 105.1
-8.6 29.3 70.0 24.7 239.0% 3) Reassay insulin levels of Rats 14 and
17 Time (min) Insulin level (.mu.U/ml), reassay Insulin level
(.mu.U/ml), original Rat #14 0 12.5 12.5 15 1020.8 1080.6 30 3018.0
3268.4 45 2590.5 3132.7 60 1714.3 2129.7 90 996.9 984.9 Rat #17 0
12.5 12.5 15 7409.9 7970.5 30 6281.1 5915.1 45 2794.8 3309.5 60
2906.4 3693.9 90 NS NS (Insulin (0.5 mg/kg), Delivery Agent (75
mg/kg))
TABLE-US-00008 TABLE 8 Oral gavage of capsules containing
containing a simple mix of insulin and delivery agent data 1)
Insulin rat time #1 #2 #3 #4 #5 #6 #7 #8 #9 #10 mean SD SE CV 0
12.5 12.5 12.5 12.5 12.5 12.5 12.5 12.5 12.5 12.5 12.5 0 0 0.0% 15
41.4 230.9 58.3 110.7 82.2 12.5 12.5 12.5 14.9 25.2 60.1 68.8 21.8
114.4% 30 12.5 49.6 12.5 12.5 12.5 12.5 12.5 12.5 12.5 12.5 16.2
11.7 3.7 72.3% 45 12.5 12.5 12.5 12.5 12.5 12.5 12.5 12.5 12.5 12.5
12.5 0 0 0.0% 60 12.5 12.5 12.5 12.5 12.5 12.5 12.5 12.5 12.5 12.5
12.5 0 0 0.0% 90 12.5 12.5 12.5 12.5 12.5 12.5 12.5 12.5 12.5 12.5
12.5 0 0 0.0% AUC.sub.0.fwdarw.90 1559 4958 1812 2598 2171 1125
1125 1125 1161 1316 1895 1189 376 62.7% 2) Glucose Change from base
line Time Rat# (min) 1 #2 #3 #4 #5 #6 #7 #8 #9 #10 mean SD SE CV 0
0 0 0 0 0 0 0 0 0 0 0 0 0 0% 15 7.7 -32.3 3.4 9.3 14.5 58.7 34 1.7
51.7 1.7 15.0 26.7 8.4 177.7% 30 -38.2 -78.5 -46.1 -42.9 -47.2 28.7
45.7 54.6 2.7 -17.9 -13.9 44.9 14.1 -322.4% 45 -13.9 -22.6 -20.8
-34.8 -38.9 20.4 42.1 44.8 -0.7 11.0 -1.3 30.1 9.5 -2247.0% 60 -3.1
41.4 -18.5 -14.3 -26.9 20.9 65.5 47.7 26.2 32.9 17.2 31.3 9.9
182.1% 90 6.2 73.1 32.6 53.4 40.1 43.1 51.7 39.6 30.6 41.2 18.4 5.8
44.6% (Insulin (0.5 mg/kg), Delivery Agent (75 mg/kg))
Example 2
1. Summary of Intravenous, Portal Vein and Subcutaneous
Experiments
[0247] a. Experiment
[0248] Intravenous, intraportal and subcutaneous dosing in rodents
were conducted to estimate the absolute bioavailability, the
absorption of insulin in the portal vein, and the bioavailability
to relative subcutaneous administration. The data are summarized in
Tables 9 to 11. The average insulin AUC.sub.0-<.infin./Dose was
0.0093 min.kg/ml from intravenous dosing. This value was assumed to
be constant in the estimates of absolute bioavailability.
TABLE-US-00009 TABLE 9 Insulin pharmacokinetics results from
intravenous (IV) administration AUC.sub.0->.infin. Dose C.sub.o
k.sub.el (min AUC.sub.0->.infin./Dose V.sub.d
C.sub.0->.infin./Dose AUC.sub.0->.infin./Dose (.mu.g/kg)
(ng/ml) (min.sup.-1) ng/ml) (min kg/ml) (ml/kg) (kg/ml) (min kg/ml)
2.5 15.79 0.89 17.74 0.0071 158 0.0063 0.0071 2.5 7.48 0.71 10.56
0.0042 334 0.0030 0.0042 2.5 13.36 0.84 15.87 0.0063 187 0.0053
0.0063 2.5 12.78 0.72 17.73 0.0071 196 0.0051 0.0071 2.5 11.26 0.72
15.75 0.0063 222 0.0045 0.0063 X .+-. SD 12.1 .+-. 2.8 0.78 .+-.
0.07 15.5 .+-. 2.6 0.0062 .+-. 0.0011 219 .+-. 68 (n = 5) 8 50.67
0.51 99.35 0.0124 157.88 0.0063 0.0124 8 52.64 0.53 99.32 0.0124
151.98 0.0066 0.0124 8 49.63 0.48 103.4 0.0129 161.19 0.0062 0.0129
X .+-. SD 51.0 .+-. 1.5 0.51 .+-. 0.03 100.7 .+-. 2.4 0.0126 .+-.
0.0003 157 .+-. 5 (n = 3) 9 46.79 0.47 99.55 0.0111 192.35 0.0052
0.0111 9 36.32 0.48 75.67 0.0084 247.80 0.0040 0.0084 9 55.30 0.45
122.89 0.0137 162.75 0.0061 0.0137 X .+-. SD 46.1 .+-. 9.5 0.47
.+-. 0.02 99.4 .+-. 23.6 0.0110 .+-. 0.0026 201 .+-. 43 (n = 3)
Average 0.0053 0.0093 SD 0.001133858 0.0033 SEM 0.000341871
0.00099
TABLE-US-00010 TABLE 10 Insulin results from portal and systemic
administrations n AUC-IP AUC-JV AUC ratio FPE (fH) 1 9.95 17.74
0.56 0.44 2 6.21 10.56 0.59 0.41 3 8.88 15.87 0.56 0.44 4 14.51
17.73 0.82 0.18 5 8.69 15.75 0.55 0.45 Average 9.65 15.53 0.62 0.38
SD 3.04 2.94 0.12 0.12
TABLE-US-00011 TABLE 11 Insulin results from subcutaneous (SC)
administration Insulin AUC.sub.0->T AUC.sub.0->T
AUC.sub.0->T/Dose dose Insulin Glucose (min T.sub.max (min (min
Study # (mg/kg) Time (uU/ml) SD SE (mg/dl) SD SE N uU/ml) (min)
C.sub.max ng/ml) kg/ml) 1 0.025 0 0.49 0.77 0.31 121 12.6 5.1 6
3640.8 15 76.1 140.0 0.005601 15 76.14 17.52 7.15 110 26.4 10.7 6
30 54.34 21.92 8.95 90 31.4 12.8 6 45 32.26 4.59 1.87 101 36.6 14.9
6 60 21.61 4.29 1.75 102 30.4 12.4 6 120 5.61 2.93 1.19 130 37.0
15.1 6 180 1.64 1.82 0.74 194 54.7 22.3 6 2 0.025 0 2.15 2.37 0.96
116 12.9 5.2 6 4234.4 15 79.5 162.9 0.006514 15 79.54 24.22 9.88
103 8.3 3.3 6 30 53.03 20.27 8.27 90 17.4 7.1 6 45 45.53 23.98 9.79
100 26.2 10.7 6 60 23.48 21.47 8.76 120 23.6 9.6 6 120 9.36 9.60
3.92 196 45.6 18.6 6 180 3.48 5.23 2.13 188 45.1 18.4 6 3 0.05 0
1.51 1.59 0.71 83 6.9 3.1 5 7230.0 15 98.8 278.1 0.005562 15 98.85
14.16 6.33 66 17.1 7.6 5 30 73.59 25.49 11.40 60 21.4 9.5 5 45
61.37 21.56 9.64 61 27.2 12.1 5 60 58.20 32.09 14.35 73 24.4 10.9 5
120 21.34 17.43 7.79 98 21.6 9.7 5 180 8.30 7.49 3.35 105 30.7 13.7
5 4 0.05 0 0.37 0.90 0.36 73 4.8 1.9 6 6407.7 15 103.4 246.5
0.004929 15 103.42 25.17 10.27 66 6.9 2.8 6 30 81.64 24.94 10.18 59
10.5 4.3 6 45 68.91 20.41 8.33 64 9.0 3.6 6 60 58.58 21.87 8.93 72
7.0 2.8 6 90 32.01 9.92 4.05 88 14.0 5.7 6 120 21.15 9.42 3.84 110
17.0 6.9 6 5 0.05 0 0 0 0 60 5.1 2.2 5 1317.0 15 34.9 50.7 0.001013
15 34.90 46.01 20.57 42 6.1 2.7 5 30 9.71 18.15 8.12 38 6.9 3.1 5
45 16.57 37.04 16.56 42 9.8 4.4 5 60 10.46 20.05 8.96 48 17.6 7.8 5
90 2.94 4.25 1.90 52 25.2 11.3 5 120 5.05 6.91 3.09 66 41.8 18.7 5
6 0.05 0 12.80 18.01 7.35 68 5.0 2.0 6 9937.5 15 191.7 382.2
0.007644 15 191.67 100.40 40.98 40 6.2 2.5 6 30 119.71 79.53 35.56
39 2.5 1.1 5 45 121.33 118.89 48.53 36 7.1 2.9 6 60 74.65 57.97
23.66 32 4.3 1.7 6 90 42.88 37.77 15.42 35 5.5 2.2 6 120 25.65
17.59 7.18 39 9.2 3.7 6 7 0.05 0 5.85 3.11 1.27 65 8.7 3.5 6 7711.1
30 126.0 296.6 0.005932 15 90.29 73.70 30.08 45 7.2 2.9 6 30 125.96
107.54 43.90 39 8.9 3.6 6 45 67.83 95.55 39.01 40 10.9 4.4 6 60
78.19 105.69 43.14 42 13.2 5.4 6 90 45.77 49.52 20.22 42 18.7 7.6 6
120 18.24 13.53 5.52 50 26.2 10.7 6 8 0.05 0 2.40 5.88 2.40 70 4.8
1.9 6 5303.3 15 131.7 204.0 0.004079 15 131.71 131.15 53.54 59 6.8
3.0 6 30 70.90 45.95 18.76 64 12.1 4.9 6 45 66.28 42.41 17.31 69
16.5 6.7 6 60 0.59 1.27 0.52 73 13.1 5.3 6 90 33.96 14.99 6.12 89
17.8 7.2 6 120 14.64 9.35 3.82 106 18.6 7.6 6 Avg 0.005159
AUC.sub.0->T/Dose (min kg/ml)
[0249] b. Results
[0250] The ratio of systemic to portal insulin was found to be
approximately 0.62 (calculated from data in Table 10). Hence, the
bioavailability in the portal vein can be calculated by dividing
the absolute bioavailability by 0.62. The portal bioavailability
provides an estimate of drug absorption from oral delivery. The
average insulin AUC.sub.0->t/Dose was 0.00516 min.kg/ml from
subcutaneous dosing. This value is used to estimate bioavailability
relative to subcutaneous. With the exception of the intravenous
data, all AUC were calculated from t=0 to the last sampling point
(i.e. AUC.sub.0->t).
[0251] In the rat model, these results from intraportal
administration suggest that the maximum absolute bioavailability of
insulin is approximately 60% from oral delivery or by any other
means of 100% GI absorption of insulin into the portal vein.
Secondly, the absolute bioavailability from SC is approximately
56%.
[0252] The estimates of bioavailability (absolute bioavailability,
portal bioavailability, relative bioavailability to subcutaneous,
and relative portal bioavailability to subcutaneous) are summarized
in FIGS. 8 to 12, and in Table 12. The estimated absolute
bioavailability from in situ dosing to the stomach and the jejunum
are shown in FIG. 8. The values of bioavailability were 5% when
dosed in the stomach and 18% when dosed in the jejunum from
microparticles containing coprocessed insulin (0.5 mg/kg) and
delivery agent (75 mg/kg).
[0253] The estimated absolute bioavailability from the tablet and
capsule formulations dosed by oral gavage in rats are shown in FIG.
9 using formulations containing 0.5 mg/kg insulin and 75 mg/kg
delivery agent. The values of bioavailability were 6% when dosed
from tablets, and 1.6% when dosed from capsules containing a simple
mix of insulin and carrier.
TABLE-US-00012 TABLE 12 Estimates of bioavailability Rel BA Rel
BA.sub.portal AUC.sub.0->T AUC.sub.0->T AUC.sub.0->T/Dose
BA.sub.portal (%) (%) (min T.sub.max C.sub.max (min (min BA (%)
[AUC.sub.0->T/Dose] [AUC.sub.0->T/Dose] uU/ml) (min) (uU/ml)
ng/ml) kg/ml) (%) (IP/JV = 0.62) sc = 0.0052 sc = 0.0052 SC 0.0052
55.67 Insulin (0.5 mg/kg) + Delivery Agent (75 mg/kg) Stomach
4778.70 15 114.50 183.80 3.68E-04 3.97 6.40 7.12 11.49 2130.75 15
72.75 81.95 1.64E-04 1.77 2.85 3.18 5.12 18958.58 45 893.86 729.18
1.46E-03 15.74 25.38 28.27 45.59 1126.65 15 12.61 43.33 8.67E-05
0.94 1.51 1.68 2.71 14425.95 45 212.73 554.84 1.11E-03 11.97 19.31
21.51 34.69 4000.73 60 140.31 153.87 3.08E-04 3.32 5.36 5.96 9.62
2793.45 15 118.66 107.44 2.15E-04 2.32 3.74 4.16 6.72 5046.23 15
158.50 194.09 3.88E-04 4.19 6.76 7.52 12.13 Mean 6657.63 28.13
215.49 256.06 5.12E-04 5.53 8.91 9.93 16.01 SD 6446.07 18.70 280.36
247.93 4.96E-04 5.35 8.63 9.61 15.50 SE 2279.03 6.61 99.12 87.65
1.75E-04 1.89 3.05 3.40 5.48 CV 96.82 66.48 130.10 96.82 9.68E+01
96.82 96.82 96.82 96.82 (%) Jejunum 13504.88 15 413.23 519.42
1.04E-03 11.21 18.08 20.14 32.48 21156.68 15 1193.25 813.72
1.63E-03 17.56 28.32 31.54 50.88 11967.90 15 669.38 460.30 9.21E-04
9.93 16.02 17.84 28.78 19689.45 15 1177.47 757.29 1.51E-03 16.34
26.36 29.36 47.35 46001.85 15 2270.55 1769.30 3.54E-03 38.18 61.59
68.59 110.62 11101.35 15 228.90 426.98 8.54E-04 9.21 14.86 16.55
26.70 39190.05 15 954.35 1507.31 3.01E-03 32.53 52.47 58.43 94.24
7234.05 15 374.88 278.23 5.56E-04 6.00 9.68 10.79 17.40 Mean
21230.78 15 910.25 816.57 1.63E-03 17.62 28.42 31.65 51.05 SD
14053.61 0 661.14 540.52 1.08E-03 11.66 18.81 20.95 33.80 SE
4968.70 0 233.75 191.10 3.82E-04 4.12 6.65 7.41 11.95 CV 66.19 0
72.63 66.19 6.62E+01 66.19 66.19 66.19 66.19 (%) Tablet 9181.20 15
468.81 353.12 7.06E-04 7.62 12.29 13.69 22.08 3692.64 15 162.11
142.02 2.84E-04 3.07 4.94 5.51 8.88 13068.89 15 700.10 502.65
1.01E-03 10.85 17.50 19.48 31.43 1125.00 0 12.50 43.27 8.65E-05
0.93 1.51 1.68 2.71 24533.57 15 1363.43 943.60 1.89E-03 20.36 32.84
36.58 59.00 4022.33 15 197.39 154.70 3.09E-04 3.34 5.38 6.00 9.67
10228.04 15 565.39 393.39 7.87E-04 8.49 13.69 15.25 24.60 3830.06
30 117.47 147.31 2.95E-04 3.18 5.13 5.71 9.21 2767.70 15 114.35
106.45 2.13E-04 2.30 3.71 4.13 6.66 1178.43 30 16.06 45.32 9.06E-05
0.98 1.58 1.76 2.83 Mean 7362.78 16.50 371.76 283.18 5.66E-04 6.11
9.86 10.98 17.71 SD 7669.89 9.01 445.61 295.00 5.90E-04 6.37 10.27
11.44 18.44 SE 2425.43 2.85 140.91 93.29 1.87E-04 2.01 3.25 3.62
5.83 CV 104.17 54.63 119.86 104.17 1.04E+02 104.17 104.17 104.17
104.17 (%) Capsule (co-dried) (N = 9) 11570.64 15 461.06 445.02
8.90E-04 9.60 15.49 17.25 27.82 3096.24 30 120.02 119.09 2.38E-04
2.57 4.15 4.62 7.45 175011.18 30 3268.40 6731.20 1.35E-02 145.27
234.30 260.93 420.86 3523.46 15 143.14 135.52 2.71E-04 2.92 4.72
5.25 8.47 4988.01 45 201.24 191.85 3.84E-04 4.14 6.68 7.44 11.99
285725.26 15 7970.50 10989.43 2.20E-02 237.16 382.52 426.00 687.10
28278.49 15 1611.31 1087.63 2.18E-03 23.47 37.86 42.16 68.00
8559.93 15 369.78 329.23 6.58E-04 7.11 11.46 12.76 20.58 18578.54
15 922.50 714.56 1.43E-03 15.42 24.87 27.70 44.68 Mean 59925.75
21.67 1674.22 2304.84 0.00 49.74 80.23 89.35 144.11 SD 100838.27
10.90 2570.42 3878.39 0.01 83.70 135.00 150.34 242.49 SE 33612.76
3.63 856.81 1292.80 0.00 27.90 45.00 50.11 80.83 CV 168.27 50.29
153.53 168.27 168.27 168.27 168.27 168.27 168.27 (%) Capsule
(co-dried) (N = 7) 11570.64 15 461.06 445.02 8.90E-04 9.60 15.49
17.25 27.82 3096.24 30 120.02 119.09 2.38E-04 2.57 4.15 4.62 7.45
3523.46 15 143.14 135.52 2.71E-04 2.92 4.72 5.25 8.47 4988.01 45
201.24 191.85 3.84E-04 4.14 6.68 7.44 11.99 28278.49 15 1611.31
1087.63 2.18E-03 23.47 37.86 42.16 68.00 8559.93 15 369.78 329.23
6.58E-04 7.11 11.46 12.76 20.58 18578.54 15 922.50 714.56 1.43E-03
15.42 24.87 27.70 44.68 Mean 11227.90 21.43 547.01 431.84 0.00 9.32
15.03 16.74 27.00 SD 9277.29 11.80 544.29 356.82 0.00 7.70 12.42
13.83 22.31 SE 3506.49 4.46 205.72 134.86 0.00 2.91 4.69 5.23 8.43
CV 82.63 55.08 99.50 82.63 82.63 82.63 82.63 82.63 82.63 (%)
Capsule (simple mix) 1558.08 15 41.37 59.93 1.20E-04 1.29 2.09 2.32
3.75 4956.66 15 230.89 190.64 3.81E-04 4.11 6.64 7.39 11.92 1812.60
15 58.34 69.72 1.39E-04 1.50 2.43 2.70 4.36 2598.35 15 110.72 99.94
2.00E-04 2.16 3.48 3.87 6.25 2171.06 15 82.24 83.50 1.67E-04 1.80
2.91 3.24 5.22 1125.00 0 12.50 43.27 8.65E-05 0.93 1.51 1.68 2.71
1125.00 0 12.50 43.27 8.65E-05 0.93 1.51 1.68 2.71 1125.00 0 12.50
43.27 8.65E-05 0.93 1.51 1.68 2.71 1161.65 15 14.94 44.68 8.94E-05
0.96 1.56 1.73 2.79 1315.97 15 25.23 50.61 1.01E-04 1.09 1.76 1.96
3.16 Mean 1894.94 10.50 60.12 72.88 1.46E-04 1.57 2.54 2.83 4.56 SD
1188.69 7.25 68.82 45.72 9.14E-05 0.99 1.59 1.77 2.86 SE 375.90
2.29 21.76 14.46 2.89E-05 0.31 0.50 0.56 0.90 CV 62.73 69.01 114.47
62.73 6.27E+01 62.73 62.73 62.73 62.73 (%) Insulin (0.25 mg/kg) +
Delivery Agent (37.5 mg/kg) Stomach 1669.50 45 48.80 64.21 2.57E-04
2.77 4.47 4.98 8.03 1249.50 15 20.80 48.06 1.92E-04 2.07 3.35 3.73
6.01 1373.25 0 41.20 52.82 2.11E-04 2.28 3.68 4.09 6.60 1192.50 0
21.50 45.87 1.83E-04 1.98 3.19 3.56 5.74 1125.00 0 12.50 43.27
1.73E-04 1.87 3.01 3.35 5.41 1125.00 0 12.50 43.27 1.73E-04 1.87
3.01 3.35 5.41 1333.22 15 26.38 51.28 2.05E-04 2.21 3.57 3.98 6.41
1125.00 0 12.50 43.27 1.73E-04 1.87 3.01 3.35 5.41 Mean 1274.12
9.38 24.52 49.00 1.96E-04 2.12 3.41 3.80 6.13 SD 186.56 15.91 13.77
7.18 2.87E-05 0.31 0.50 0.56 0.90 SE 65.96 5.63 4.87 2.54 1.01E-05
0.11 0.18 0.20 0.32 CV 14.64 169.71 56.16 14.64 1.46E+01 14.64
14.64 14.64 14.64 (%) Jejunum 8260.50 15 428.50 317.71 1.27E-03
13.71 22.12 24.63 39.73 10947.00 15 532.40 421.04 1.68E-03 18.17
29.31 32.64 52.65 5229.00 15 232.60 201.12 8.04E-04 8.68 14.00
15.59 25.15 1125.00 0 12.50 43.27 1.73E-04 1.87 3.01 3.35 5.41
1910.94 15 61.95 73.50 2.94E-04 3.17 5.12 5.70 9.19 3735.93 15
186.56 143.69 5.75E-04 6.20 10.00 11.14 17.97 2157.20 15 79.60
82.97 3.32E-04 3.58 5.78 6.43 10.38 3897.03 15 160.54 149.89
6.00E-04 6.47 10.43 11.62 18.74 Mean 4657.82 13.13 211.83 179.15
7.17E-04 7.73 12.47 13.89 22.40 SD 3392.58 5.30 182.48 130.48
5.22E-04 5.63 9.08 10.12 16.32 SE 1199.46 1.88 64.52 46.13 1.85E-04
1.99 3.21 3.58 5.77 CV 72.84 40.41 86.14 72.84 7.28E+01 72.84 72.84
72.84 72.84 (%)
Example 3
Insulin and 4-CNAB Stability in Simulated Gastric Fluid
[0254] The stability of insulin in simulated gastric fluid (SGF)
was evaluated in the presence and absence of 4-CNAB. Solutions were
prepared containing insulin (1 mg/ml) with and without monosodium
4-CNAB (1 mg/ml).
[0255] The SGF was prepared with and without pepsin, a gastric
enzyme. SGF pH 1.2 was prepared as per the USP NF 26 guidelines. 2
g sodium chloride and 3.2 g of pepsin were weighed and added to a
suitable container, and deionized water was added to reach one
liter in volume. If necessary, the pH was adjusted to 1.2 by
addition of concentrated HCl or NaOH. A second SGF solution
omitting the pepsin was also prepared.
[0256] Four 50 ml samples of SGF (two with pepsin and two without)
were placed into a jacketed vessel connected to a circulating water
bath set at 37.degree. C. The solutions were stirred with magnetic
stir bars for ten minutes to allow the solutions to reach
37.degree. C. and reach thermal equilibrium. 50 mg of 4-CNAB was
added to one of the samples containing pepsin and one of the
samples without pepsin, and the solutions were stirred for a few
minutes to allow the 4-CNAB to dissolve. 50 mg of insulin was added
to the each of the samples. After dissolution of the insulin,
samples of the solutions were taken at pre-determined time
intervals, filtered, and immediately assayed by HPLC for insulin
and 4-CNAB content. The first sample withdrawn after all the
insulin was dissolved was considered to have been drawn at time
zero (0). The results are shown in table 13.
TABLE-US-00013 TABLE 13 Insulin Insulin without 4- without 4- CNAB
CNAB and with and Insulin with 4-CNAB Enzymes without Insulin with
4-CNAB and Enzymes (pepsin) (pepsin) Enzymes and no Enzymes Insulin
% 4-CNAB % Insulin % Insulin % Insulin % 4-CNAB % of of of of of of
Time theoretical theoretical theoretical theoretical theoretical
theoretical 0 minutes 3.0 105.6 3.0 99.8 100.8 95.3 10 minutes
100.4 103.9 95.7 20 minutes 100.6 103.8 99.6 30 minutes 100.1 105.7
99.2 1 h 99.4 97.8 94.8 2 h 99.0 102.2 95.8 24 h 0 0 91.2
[0257] The term "% of theoretical" as used herein, means the
percent of the concentration (mg/mL) of withdrawn solution at the
time-point the sample was taken as compared to the theoretical
concentration (mg/mL) of the measuring component for experiment.
The standard of deviation for the HPLC analysis is .+-.5%. These
results show that insulin is unstable in SGF containing pepsin,
since only 3.0% of the insulin remained at the first sampling point
(97% of the insulin was degraded), while insulin is stable at least
up to 2 hours in SGF without pepsin.
Example 4
Stability of Insulin in Simulated Intestinal Fluid
[0258] The stability of insulin in simulated intestinal fluid (SIF)
was evaluated in the presence and absence of 4-CNAB.
[0259] The SIF solutions were prepared with and without pancreatic
enzyme. SIF pH 7.5 was prepared as per the USP NF 26 guidelines.
SIF was prepared by addition of 6.8 g monobasic potassium phosphate
and 10 g of pancreatin into a suitable vessel, and deionized water
was added to reach a total volume of one liter. If necessary, the
pH was adjusted to 7.5 by addition of 0.2 N sodium hydroxide. A
second SIF solution omitting the pancreatin, an intestinal enzyme,
was also prepared.
[0260] Four 50 ml samples of SIF (two with pancreatin and two
without) were placed into a jacketed vessel connected to a
circulating water bath set at 37.degree. C. The solutions were
stirred with magnetic stir bars for ten minutes to allow the
solutions to reach 37.degree. C. and reach thermal equilibrium. 50
mg of 4-CNAB was added to one of the samples containing pepsin and
one of the samples without pepsin, and the solutions were stirred
for a few minutes to allow the 4-CNAB to dissolve. 50 mg of insulin
was added to the each of the samples. After dissolution of the
insulin, samples of the solutions were taken at pre-determined time
intervals, and immediately assayed by HPLC for insulin and 4-CNAB
content. The results are shown in table 14.
TABLE-US-00014 TABLE 14 Insulin without 4- Insulin CNAB and Insulin
with 4-CNAB without 4- with and Enzymes CNAB or Enzymes Insulin
with 4-CNAB (pancreatin) Enzymes (pancreatin) and no Enzymes
Insulin % 4-CNAB % Insulin % 4-CNAB % Insulin % 4-CNAB % of of of
of of of Time theoretical theoretical theoretical theoretical
theoretical theoretical 0 minutes 58.9 98.6 92.9 66.9 100.2 103.9 2
minutes 26.9 98.9 -- 55.2 102.1 105.4 4 minutes 19.3 99.0 -- 45.1
106.3 111.5 6 minutes 5.4 98.8 -- 34.5 103.1 106.0 8 minutes 3.1
98.6 -- 24.5 101.7 101.8 10 minutes 2.5 98.2 92.5 15.1 101.3 103.4
15 minutes 92.7 100.9 104.0 30 minutes 94.7 100.6 104.0 45 minutes
-- 99.1 102.3 1 hour 92.7 95.9 100.0 2 hours 94.6 -- -- 24 hours
100.4 105.6 111.3
[0261] These results show that insulin is stable in SIF without
pancreatin and degrades in presence of the enzyme. Insulin is more
stable in SIF with and without enzyme than in SGF with and without
enzyme. At the first sampling time point (0 minuts) only 3.0%
insulin remained in SGF with enzymes while 58.9% and 66.9% insulin
remained in SIF.
Example 5
Effect of Formulation on Insulin Absorption and Action
[0262] Six formulations containing insulin shown in Table 15 were
prepared as follows.
TABLE-US-00015 TABLE 15 ID Formulation 1 Fast Disintegrating
Tablets-1. Insulin, 4-CNAB, 0.4% w/w povidone, 10% w/w Polyplasdone
XL, 50.7% w/w Emcocel HD90, 1% w/w SLS and 1% w/w Magnesium
Stearate 2 Fast Disintegrating Tablets-2. Insulin, 4-CNAB, 0.4% w/w
povidone 10% w/w Polyplasdone XL, 50.2% w/w Prosolv HD90, 1% w/w
SLS and 1% w/w Magnesium Stearate 3 Tablets with Emcompress.
Insulin, 4-CNAB, 0.4% w/w povidone, ~29.1% w/w Emcompress, 1% w/w
SLS and 1% w/w Magnesium Stearate 4 Tablets with Mg Stearate only.
Insulin, 4-CNAB, 0.4% w/w povidone, and 1% w/w Magnesium Stearate 5
Tablets with Anhydrous Emcompress. Insulin, 4-CNAB, 0.4% w/w
povidone, ~29.1% w/w Anhydrous Emcompress, 1% w/w SLS and 1% w/w
Magnesium Stearate 6 Co-dried capsules. Insulin and 4-CNAB were
dissolved in water. The solution was co-dried and the resulting
powder was filled into hard gelatin capsules.
[0263] Polyplasdone XL, is available from International Specialty
Products, Wilmington Del.; Emcocel HD90, Prosolv HD90, Emcompress
and Anhydrous Emcompress is available from JRS Pharma, Patterson,
N.Y.
[0264] The formulations were fed to rhesus monkeys in doses
containing 100 mg/kg of 4-CNAB and 13 U/kg insulin. Groups of four
rhesus monkeys, two males and two females, were fasted for at least
12 hrs prior to dosing and up to 4 hrs after dosing. Water was
withheld approximately 1 hr before dosing and up to 2 hrs after
dosing after which it was permitted ad libitum. The dosing was
followed by a 5 ml water flush. Blood samples (approximately 2 ml
each) were collected by venipuncture at 15 minutes before dosing
and at 5, 10, 15, 20, 30, 45 minutes and 1, 1.5, 2, 3, 4 hr after
dosing. Each blood sample was divided into two portions. One
portion was allowed to clot at room temperature and centrifuged at
2-8.degree. C. for 10 minutes at 3000 rpm. The serum obtained was
aliquoted into two portions and stored at -70.degree. C. until
shipment. One sample was shipped to Emisphere on dry ice for
insulin analysis by ELISA while the other was retained by the CRO
for serum glucose analysis. The second portion of the blood was
kept on wet ice for up to 30 minutes and centrifuged at 2-8.degree.
C. for 10 minutes at 3000 rpm. The plasma obtained was shipped to
Emisphere on dry ice for analysis of 4-CNAB content by HPLC. Each
formulation was administered to 4 rhesus monkeys, except
formulation 1, which was administered to 8 rhesus monkeys. Blood
samples were taken at predetermined intervals as described above
and assayed for insulin and glucose levels. The results are shown
in table 16 and in FIGS. 17 and 18.
TABLE-US-00016 TABLE 16 Serum Glucose Onset of t.sub.min Disint.
glucose Insulin (Cmin, % change Duration of Action Form.ID Time
lowering t.sub.max from baseline .+-. SE) (Glucose Level) 1 35 sec
4 min 30 min 120 min 240 min + (-33.57 .+-. 8.25%) (-21.87 .+-.
6.64%) 2 23 sec 8 min 10 min 60 min 240 min + (-25.6 .+-. 13.91%)
(-26.17 .+-. 7.39%) 3 6 min 7 min 60 min 60 min 240 min + 43 sec
(-50.68 .+-. 4.41%) (-33.96 .+-. 3.36%) 4 8 min 2 min 15 min 45 min
240 min 40 sec (-46.27 .+-. 7.78%) (-0.80 .+-. 13.98%) 5 7 min 6
min 20 min 60 min 240 min + 6 sec (-58.13 .+-. 3.89%) (-10.14 .+-.
0%) 6 9 min 15 min 45 min 90 min (-26.96 .+-. 13.13%) (18.43 .+-.
23.84%)
[0265] Disintegration time was determined in water at
37.+-.2.degree. C. using the method described in USP <701>.
Multiple tubes containing water are placed in a basket-rack
assembly immersed in a water bath maintained at 37.+-.2.degree. C.
The basket-rack assembly raises and lowers the tubes at a constant
frequency. The tablets are placed in the tubes and are periodically
examined to determine if they have disintegrated completely. Each
tablet is tested in six different tubes. If 1 or 2 tablets fails to
consistently disintegrate, the procedure is repeated on additional
tablets. The average maximum concentration of insulin (C.sub.max)
was determined for each group based upon the serum levels of
insulin measured as described above. If the blood glucose levels in
the primates falls to very low levels (<1 mmol/L) during the
experiment they are administered dextrose in order to bring the
blood glucose up to a safe level. The average C.sub.max for each
group, as well as the number of rhesus monkeys rescued, is shown in
table 17.
TABLE-US-00017 TABLE 17 Formulation Insulin C.sub.max No. of
primates No. of primates ID (.mu.U/ml) dosed rescued 1 0.6585 .+-.
0.6585 8 0 2 3.99 .+-. 3.99 4 0 3 70.81 .+-. 43.22 4 2 4 51.65 .+-.
30.69 4 1 5 60.46 .+-. 34.56 4 3 6 31.75 .+-. 31.54 4 1
Example 6
Preparation of Enteric Coated Tablets
[0266] Capsules were manufactured by encapsulating 300 mg of a
formulation including 150 units insulin, 200 mg 4-CNAB, 0.4% w/w
povidone, .about.29.1% w/w Emcompress, 1% w/w SLS, and 1% w/w
magnesium stearate into size 2 white opaque capsules. The capsules
were first coated with a subcoat consisting of Opadry clear for a
weight gain of 5% followed by an enteric coat of 20% weight gain
for a total weight gain on the capsules of 25%.
[0267] Tablets were manufactured by pressing 300 mg of the
formulation described above into tablets. An 10% weight gain
enteric coat was applied. The formulations for the subcoats and
enteric coats are shown in table 18 below.
TABLE-US-00018 TABLE 18 Tablets Capsules Ingredients % w/w % w/w
SUBCOAT Opadry Clear NA 8.0 Milli Q Water NA 92.0 Total NA 100.0
ENTERIC COAT Eudragit L3 0D55 49.4 49.4 Talc 3.7 3.7 Triethyl
Citrate 1.5 1.5 Milli Q Water 45.4 45.4 Total 100.0 100.0
[0268] Opadry.TM. Clear is available from Colorcon, of West Point,
Pa.
[0269] Milli Q Water is highly purified water and is available from
Millipore of Billerica, Mass.
[0270] Eudragit L30D55 is available from Degussa AG, Parsippany,
N.J.
[0271] To verify the effectiveness of the enteric coat, the coated
capsules and tablets were placed in 0.1 N HCl for two hours or pH
6.8 phosphate buffer for one hour. The coated capsules and tablets
did not dissolve in the 0.1 N HCl, but did dissolve in the pH 6.8
phosphate buffer.
Example 7
SNAC Micro Beads Coated With Heparin
[0272] 5 g of SNAC and 0.5 g of magnesium stearate were mixed. 0.02
g of the mixed powder was fed into a die. Small beads of SNAC and
magnesium stearate were made at 1200 PSI bar pressure The beads had
a round/ball shape size of about 0.2 mm to about 2.0 mm. The SNAC
beads were then coated with 2.5 g of heparin, in liquid form, by a
rotary method and dried under vacuum oven at 40.degree. C. for 10
hours.
Example 8
Micronized SNAD with Heparin
[0273] SNAD was screened through a 35 mesh Tyler standard sieve.
The SNAD was milled with a Glen Mills, Model S100 centrifugal ball
mill (Clifton, N.J.) equipped with a 250 mL stainless steel
grinding jar and 30 mm (440c) diameter stainless steel balls was
used. The process parameters investigated were (1) number of balls
used, (2) duration of milling, (3) milling speed, and (4) milling
jar total charge. A Malvern Mastersizer 2000 equipped with a
Scirocco 2000 dry accessory was used for particle size
determination. A Kratos XRD 6000 (version 4.1) X-ray powder
diffractometer scanning over the 20 range 5-40.degree. 20 was used
for monitoring crystallinity changes. The diverging, scattering,
and receiving slits were 10.degree., 10, and 0.3 mm respectively. A
Brinkmann 737 KF coulometer was used for moisture content
determination while a Quantachrome Nova 3000 Series Surface Area
Analyzer was used for specific surface area determination.
[0274] The results indicated that the particle size distribution of
pre-screened SNAD was d(0.1)=1.6 .mu.m, d(0.5)=10.5 .mu.m, and
d(0.9)=314.9 .mu.m. The data obtained using different numbers of
balls ranging from 1 to 5 indicated that the optimum number of
balls for the charge used was 2. The use of 2 balls yielded the
particle size d(0.1)=1.1 .mu.m, d(0.5)=12.0 m, and d(0.9)=154.3
.mu.m.
[0275] An evaluation of the effect of milling time for a fixed
number of balls and charge indicated that a milling time of 120
minutes was optimum resulting in the particle size distribution,
d(0.1)=2.0 .mu.m, d(0.5)=15.4, and d(0.9)=62.9 .mu.m.
[0276] An evaluation of the milling speeds 100, 300, and 500 rpm
indicated that optimum milling was obtained at 300 rpm. This speed
yielded the particle size distribution, d(0.9)=62.9 m compared to
unmilled SNAD d(0.9)=314.9 .mu.m.
[0277] A charge of 37 mL of the 250 mL milling jar provided better
milling compared to 75 and 112 mL. The powder X-ray diffraction
analysis indicated that milling did not result in crystallinity
changes for SNAD. The Karl Fischer moisture content determination
indicated no significant changes in moisture content.
[0278] The SNAD was then mixed with heparin.
Example 9
Micronized SNAC with Micronized Heparin
[0279] SNAC and heparin were micronized separately by the procedure
described in Example 8 with 2 balls at 200 rpm for 120 minutes and
then mixed together. The micronized SNAC had a d(0.5) of 7.574
.mu.m SNAC/heparin capsules having the formulations shown in table
19 below were prepared by hand packing them into hard gelatin
capsules.
TABLE-US-00019 TABLE 19 Formulation (mg/capsule) Ingredient A B
Micronized SNAC 125 125 Micronized Heparin USP 158 158 (30,000 U)
Propylene Glycol 105 105 Monocaprylate.sup.1 Sodium lauryl sulfate
7 7 PEG 300.sup.2 305 270 Water -- 35 Total 700 700
[0280] The heparin, SNAC, and sodium lauryl sulfate were mixed.
Separately, the PEG 300, propylene glycol monocaprylate, and water
(for formulation B) were mixed. 50% of the liquid PEG 300/propylene
glycol monocaprylate mixture was transferred to a mortar. The
heparin, SNAC, and sodium lauryl sulfate blended powder was added
little by little and triturated with the liquid in the mortar and
pestle. The capsules were then packed with the resulting mixture.
.sup.1--Propylene glycol monocaprylate is available as Capmul.TM.
PG 8 from Abitec Corporation of Columbus, Ohio.sup.2--PEG 300 is
available as Carbowax.TM. 300 from Dow Chemical Co. of Midland,
Mich.
Example 10
Micronized SNAC/Heparin
[0281] Heparin (118.5 mg/dose (22,500 rpm)) and SNAC (125 mg/dose)
were dry mixed, screened through a 35 mesh screen, and milled for
about 4 minutes with a ball mill. The mixture was packed into
capsules (Capsugel Size 1 capsules (Greenwood, S.C.)).
[0282] The capsules were administered to rhesus monkeys (2 capsules
per monkey) by the following procedure. Rhesus monkeys weighing
between 3.5-5.0 kg were fasted overnight before the experiments and
food was returned about 2 hours after dosing. Water was withheld
from 30 minutes prior to dosing until 30 minutes after dosing,
except for those quantities used for dosing. Each dosage form was
delivered to the rear of the mouth using a pill gun. After release
of the dosage form, 5 ml of reverse osmosis water was administered
into the oral cavity to facilitate swallowing. Following delivery,
the oral cavity was inspected to ensure that the capsule was
swallowed. Antifactor Xa from blood samples was measured over 6
hours.
[0283] The results are shown in FIG. 19.
Example 11
Micronized SNAC/Heparin
[0284] Capsules containing micronized SNAC/heparin as shown in
table 20 below were prepared as follows.
[0285] A solution of heparin and SNAC was prepared as follows. The
required amounts of heparin and SNAC were weighed out and water,
which was previously adjusted to a pH of about 8 with sodium
hydroxide, was added. The pH of the resulting solution was in the
range of about 7.3-7.5. The solution pH was adjusted to a pH of
about 8 with sodium hydroxide. The solution was then dried in a
RotoVap apparatus at 50.degree. C. under vacuum. The evaporating
was done using the program outlined below.
[0286] 1. Immediate reduction of vacuum from 760 torr to 200
torr
[0287] 2. Reduction of vacuum pressure from 200 to 100 torr in 2
minutes
[0288] 3. Reduction of vacuum pressure from 100 to 50 torr in 2
minutes
[0289] 4. Reduction of vacuum pressure from 50 to 25 torr in 4
minutes
[0290] 5. Reduction of vacuum pressure from 25 to 15 torr in 4
minutes
[0291] 6. Reduction of vacuum pressure from 15 to 10 torr in 2
minutes
[0292] 7. Evaporating at 10.+-.2 torr and 70 rpm in 30 minutes
[0293] 8. Switch to 50 rpm manually and continue with evaporating
for 4 hours
[0294] The sample was vacuum dried overnight. The resulting powder
was then micronized and filled into capsules to give the desired
dose.
TABLE-US-00020 TABLE 20 Formulation (mg/capsule) Ingredient A B C
Micronized SNAC 198 173 210.6 Micronized 72 63 78 Heparin USP
(13104 USP (11466 USP (14196 USP heparin units) heparin units)
heparin units) Total 270 236 288.6
Prophetic Example 12
[0295] Micronized 5-CNAC disodium and tablets of salmon calcitonin
plus micronized 5-CNAC disodium may be prepared in accordance with
the present invention as follows:
Preparation of Micronized 5-CNAC Disodium
[0296] Coarse 5-CNAC disodium, which is to be micronized, is added
to a jet mill (Air Jet Mill GemT.RTM., Copley Scientific, Ltd.,
Nottingham, UK) using a 80 ceramic pan cake jet mill, 8 cm
diameter, 6 bar N2, 0.5 mm nozzles with manual feed of about 700
g/h. The coarse 5-CNAC disodium is jet milled and periodically
sampled under microscope with reference ruler measurements to
identify when the average desired micronized particle size is
obtained. Three different batches are ground to create 6 .mu.m, 35
.mu.m, and 46 .mu.m batches. Individual sieving of the separate
micronized batches is then done by using a conical sieve mill
(Quadro Comil, Quadro Engineering Incorporated 613 Colby Drive,
Waterloo, Ontario, Canada N2V 1A1) with a U10, 813 .mu.m conical
sieve, round beater, operating at 1500 upm with throughput of about
150 kg/h.
Formulation I
TABLE-US-00021 [0297] Salmon Calcitonin Formulation with 5-CNAC
Disodium of Different Particle Size Ingredient Amount (mg) Percent
(%) Salmon Calcitonin 1 0.25 Micronized 5-CNAC 228 57 Disodium
Avicel PH 102 .RTM. 147 36.75 Crospovidone, NF 20 5 Magnesium
stearate 4 1 Total 400 100
Preparation of Formulation 1
[0298] Three different batches of tablets are prepared using the
three different batches of micronized 5-CNAC disodium, one tablet
batch having an average 5-CNAC disodium particle size of 46 microns
(Batch A), a second tablet batch having an average 5-CNAC disodium
particle size of 6 microns (Batch B), and a third tablet batch
having an average 5-CNAC disodium particle size of 35 microns
(Batch C).
[0299] 0.50 g of salmon calcitonin, pre-screened through a 40 mesh
screen, 57 g of micronized 5-CNAC disodium salt, screened through a
35 mesh screen, and 10 g of Polyplasdone XL (crospovidone, NF,
International Specialty Products, 1361 Alps Road, Wayne, N.J.,
07470, USA) is combined in a 500 mL jar and is mixed using a
Turbula mixer for 100 revolutions at a speed of 46 RPM. An
additional 57 g of micronized 5-CNAC disodium salt, screened
through a 35 mesh screen, and 36.75 g of Avicel PH 102.RTM. is
added to the jar and mixed for 500 revolutions at a speed of 46
RPM. A further 36.75 g of Avicel PH 102.RTM. is added to the jar
and is mixed for an additional 100 revolutions at a speed of 46
RPM. 4.0 g of magnesium stearate is screened into the jar using a
35 mesh screen and is blended for 1 minute at a speed of 46 RPM.
The final blend is compressed into tablets using a Manesty B3B
tablet press. The tablet weight is approximately 400 mg.
[0300] The bioavailability of the tablets created in this example
may be tested as described in Example 13.
Prophetic Example 13
Primate Administration
[0301] The tablets are prepared as in Example 12 using three
different batches of micronized 5-CNAC disodium, one tablet batch
having an average 5-CNAC disodium particle size of 46 microns
(Batch A), a second tablet batch having an average 5-CNAC disodium
particle size of 6 microns (Batch B), and a third tablet batch
having an average 5-CNAC disodium particle size of 35 microns
(Batch C). Each tablet contains 200 mg 5-CNAC disodium and 1 mg
salmon calcitonin. The tablets prepared from each of the three
different batches are administered to the same four Rhesus monkeys
separately on different days as follows:
[0302] The Rhesus monkeys fast overnight prior to dosing and are
restrained in chairs fully conscious, for the duration of the study
period. One tablet from Batch A or Batch B or Batch C is
administered to each monkey via a gavage tube followed by 10 mL of
water.
[0303] Rhesus monkey blood samples are collected immediately before
administration and at 0.25, 0.5, 0.75, 1, 1.5, 2, 3, 4, 5, and 6
hours after administration. A tablet from each of the remaining two
tablet batches is dosed and blood samples are collected in a
similar manner but on a separate day for each of the remaining
tablet batches. Resulting plasma salmon calcitonin for each dose
and for each monkey is determined by radioimmunoassay. For each
monkey, the primate plasma salmon calcitonin (SCt) for one batch
and one time period, mean plasma SCt concentrations for all monkeys
for one batch and one time period, Standard Deviation (SD) of
plasma SCt concentrations for one batch and one time period, and
Standard Error of the Mean (SEM) for plasma SCt concentrations for
all monkeys for one batch and one time period are calculated. The
prophetic results are shown in the tables below.
TABLE-US-00022 BATCH A: AVERAGE 5-CNAC PARTICLE SIZE 46 MICROMETERS
Salmon Calcitonin (SCt) Plasma Concentrations [pg/mL] (Single Oral
Tablet (200 mg 5-CNAC + 1 mg SCt) to the Rhesus Monkey) Animal Time
[hours] no. 0 0.25 0.50 0.75 1 1.5 2 3 4 5 6 1 0.0 17.8 91.7 279.7
449.2 278.8 48.0 10.5 5.3 3.3 0.0 2 0.0 117.4 535.0 430.8 981.4
1718.0 2396.4 719.5 253.6 102.1 62.9 3 0.0 113.9 754.5 1502.0
2351.0 2066.0 2684.4 1310.0 649.6 280.6 156.5 4 0.0 46.0 127.0
425.5 765.8 1102.0 1599.0 1022.0 419.3 87.0 23.4 Mean 0.0 73.8
377.1 659.5 1136.9 1291.2 1682.0 765.5 332.0 118.3 60.7 SD 0.0 49.7
322.2 566.0 838.4 783.8 1182.1 558.1 271.6 116.6 68.9 SEM 0.0 24.9
161.1 283.0 419.2 391.9 591.0 279.0 135.8 58.3 34.5 Lower Limit of
Quantification (LLOQ) = 2.5 pg/mL, concentrations below LLOQ were
set to zero for Table 1
TABLE-US-00023 BATCH B: AVERAGE 5-CNAC PARTICLE SIZE 6 MICROMETERS
Salmon Calcitonin (SCt) Plasma Concentrations [pg/mL] (Single Oral
Tablet (200 mg 5-CNAC + 1 mg SCt) to the Rhesus Monkey) Animal Time
[hours] no. 0 0.25 0.50 0.75 1 1.5 2 3 4 5 6 1 0.0 265.6 315.8
245.6 357.2 1927.0 3010.0 863.2 139.4 48.5 20.8 2 0.0 607.0 777.0
1336.0 1602.0 4146.0 7521.0 2681.0 420.8 73.9 43.2 3 0.0 80.9 225.5
325.6 655.6 1478.0 3979.0 2775.0 520.2 91.5 41.3 4 0.0 286.4 155.3
237.7 241.0 269.7 294.2 321.0 179.8 67.5 13.6 Mean 0.0 310.0 368.4
536.2 714.0 1955.2 3701.1 1660.1 315.1 70.4 29.7 SD 0.0 218.5 280.2
534.7 617.2 1619.6 2986.3 1253.5 184.8 17.8 14.8 SEM 0.0 109.2
140.1 267.3 308.6 809.8 1493.1 626.7 92.4 8.9 7.4 Lower Limit of
Quantification (LLOQ) = 2.5 pg/mL, concentrations below LLOQ were
set to zero for Table 2
TABLE-US-00024 BATCH C: AVERAGE 5-CNAC PARTICLE SIZE 35 MICROMETERS
Salmon Calcitonin (SCt) Plasma Concentrations [pg/mL] (Single Oral
Tablet (200 mg 5-CNAC + 1 mg SCt) to the Rhesus Monkey) Animal Time
[hours] no. 0 0.25 0.50 0.75 1 1.5 2 3 4 5 6 1 0.0 36.1 94.7 428.0
739.4 2568.0 4025.0 1348.0 499.6 218.4 98.1 2 0.0 10.9 55.0 168.9
248.2 507.3 654.0 434.8 177.3 68.8 38.9 3 0.0 172.3 336.6 409.5
584.9 1487.0 2087.0 1479.0 162.0 52.0 17.2 4 0.0 7.9 46.9 208.1
390.1 1237.0 2347.0 1342.0 192.3 42.3 19.2 Mean 0.0 56.8 133.3
303.6 490.7 1449.8 2278.3 1151.0 257.8 95.4 43.4 SD 0.0 78.0 137.1
134.1 215.8 853.5 1382.1 481.6 161.7 82.7 37.8 SEM 0.0 39.0 68.6
67.1 107.9 426.7 691.1 240.8 80.8 41.4 18.9 Lower Limit of
Quantification (LLOQ) = 2.5 pg/mL, concentrations below LLOQ were
set to zero for Table 3
[0304] The compositions according to the instant invention allow
considerably improved oral bioavailability of active agent. The
improved bioavailability results in high in vivo concentrations of
active agent, particularly calcitonin, being achieved via oral
delivery, and in correlation to the particle sizes of 5-CNAC in the
oral formulations of the examples.
[0305] The present invention is not to be limited in scope by the
specific embodiments described herein. Indeed, various
modifications of the invention in addition to those described
herein will become apparent to those skilled in the art from the
foregoing description and the accompanying figures. Such
modifications are intended to fall within the scope of the appended
claims.
[0306] Patents, patent applications, publications, product
descriptions, and protocols are cited throughout this application,
the disclosures of which are incorporated herein by reference in
their entireties for all purposes.
* * * * *